GFP antibody
Principal name
GFP antibody
Alternative names for GFP antibody
Green fluorescent protein, GFP-Tag
SwissProt ID
P42212 (Aeqvi)
Gene ID
3020 (H3F3A)
Available reactivities
Available hosts
Available applications
Western blot / Immunoblot (WB), Frozen Sections (C), Immunoprecipitation (IP), Immunocytochemistry/Immunofluorescence (ICC/IF), Enzyme Immunoassay (E), Paraffin Sections (P), ChIP assay (ChIP), In Situ Hybridization (ISH), Immunoelectron Microscopy (EM), Dot blot (Dot), Flow Cytometry (F), Immunohistochemistry (IHC)
Background of GFP antibody
Green fluorescence protein (GFP) is a 27 KDa protein derived from the bioluminiscent jellyfish Aquorea victoria, emiting green light (509 nm) when excited (excitation by Blue or UV light, absorption peak at 395 nm).
GFP is a useful tool in cell biology research, as its intrinsic fluorescence can be visualized in living cells. Light-stimulated GFP fluorescence is species-independent and a fluorescence has been reported from many different types of GFP-expressing hosts, including microbes, invertebrates, vertebrates and plants. No exogenous substrates and cofactors are required for the fluorescence of GFP, since GFP autocatalytically forms a fluorescent pigment from natural amino acids present in the nascent protein.
GFP fluorescence is stable under fixation conditions and suitable for a variety of applications. GFP is widely used as a reporter (tag) for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without any further staining. Other applications of GFP include measurement of distance between proteins through fluorescence energy transfer (FRET) protocols.
To increase a fluorescence intensity of GFP, chromophore mutations have been created. The Enhanced GFP has a fluorescence 35 times more intense than the wt-GFP. Mutagenesis of GFP has produced also many mutants (e.g. Yellow Fluorescent Protein, Cyan Fluorescent Protein) with warying spectral properties. Antibodies raised against full-length GFP variants should also detect other variants of the protein.
General readings
Schoch KG et al. A subset of mouse tracheal epithelial basal cells generates large colonies in vitro. Am J Physiol Lung Cell Mol Physiol 286:L631-42 (2004). PubMed PMID: 12959927
Karabay A. et al Axonal growth is sensitive to the levels of katanin, a protein that severs microtubules. The Journal of Neuroscience 24:5778-5788 (2004).Gongidi V et al. SPARC-like 1 Regulates the Terminal Phase of Radial Glia-Guided Migration in the Cerebral Cortex. Neuron 41:57-69 (2004). PubMed PMID: 14715135
Fischer AC et al. Successful transgene expression with serial doses of aerosolized rAAV2 vectors in rhesus macaques. Mol Ther 8:918-26 (2003). PubMed PMID: 14664794
Wong JM et al. Subnuclear shuttling of human telomerase induced by transformation and DNA damage. Nat Cell Biol 4:731-6 (2002). PubMed PMID: 12198499
Zhang QX et al. Identification of structurally important domains of lipid phosphate phosphatase-1: implications for its sites of action. Biochem J 345 Pt 2:181-4 (2000). PubMed PMID: 10620492
Mesaeli N et al. Calreticulin is essential for cardiac development. J Cell Biol 144:857-68 (1999). PubMed PMID: 10085286
McCabe JB & Berthiaume LG Functional roles for fatty acylated amino-terminal domains in subcellular localization. Mol Biol Cell 10:3771-86 (1999). PubMed PMID: 10564270
Jasinska R et al. Lipid phosphate phosphohydrolase-1 degrades exogenous glycerolipid and sphingolipid phosphate esters. Biochem J 340 ( Pt 3):677-86 (1999). PubMed PMID: 10359651
1. Reinhardt, R. L., Hong, S., Kang, S.-J., Wang, Z.-e., and Locksley, R. M. (2006) Visualization of IL-12/23p40 In Vivo Reveals Immunostimulatory Dendritic Cell Migrants that Promote Th1 Differentiation, 177, 1618-1627.
Figure 1. Confocal microscopy images of COS-7 cells transfected with expression constructs encoding membrane-tethered EGFP (membrane-EGFP; top) or nuclear Polycomb 2-EYFP fusion protein (Pc2-EYFP; bottom). The natural fluorescence of the produced proteins is shown in the green channel (left), the anti-GFP antibody signal was detected in the red channel (right). The system was carefully tested for overlap of these two optical channels and images were scanned separately in sequential scanning mode. The blue nuclear stain is also shown.
Figure 2. Immunoprecipitation of GFP-NLS from HEK293 cells using anti-GFP antibody. HEK293 cells were transfected with expression construct encoding GFP-NLS protein. Twenty hours post transfection cells were lysed in non-denaturating conditions (Lysis buffer: 20 mM Tris, pH 7.5, 100 mM NaCl, 0.5% Triton X-100, inhibitors of proteases). Aliquots of cell lysate were immunoprecipitated using a rabbit anti-GFP antibody (lane 2) or a pre-immune rabbit serum (lane 3). Immunoprecipitates together with a sample of the cell lysate (lane 1) were separated on SDS-PAGE polyacrylamide gel and immunoblotted with the anti-GFP antibody. The positions of molecular weight markers in kDa are indicated at the left.
Immunofluorescence: GFP Antibody [NB100-1770] - anti-GFP antibody at a 1:1,000 dilution detects tau-GFP in cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) in Drosophila melanogaster late stage embryonic central nerve cells.
Immunofluorescence: GFP Antibody [NB100-1770] - Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus in Mus musculus (coronal view, 20X magnification). Briefly, Sf-1:Cre mice (Jackson Mouse Laboratories) were crossed to the Z/EG reporter line.
Western Blot: GFP Antibody [NB100-1770] - Lane 1 shows detection of a 33 kDa band corresponding to a GFP containing recombinant protein (arrowhead) expressed in HeLa cells. Lane 2 shows no staining of a mock transfected HeLa cell lysate.
Immunofluorescence: GFP Antibody [FITC] [NB100-1771] Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus in Mus musculus (coronal view, 20X magnification). Briefly, Sf-1:Cre mice were crossed to the Z/EG reporter line. Brains were harvested following cardiac perfusion with 4%PFA/PBS with o/n fixation, and subsequently transferred to a 30% sucrose solution. Brains were then frozen in OCT freezing medium and cryostat sectioned at 40 microns. Goat anti-GFP was used at 1:500 dilution in free floating imunnohistochemistry to detect eGFP. Fluorchrome conjugated anti-goat IgG secondary antibody was used for detection at 1:500. Sections were counterstained with DAPI.
Immunofluorescence: GFP Antibody [FITC] [NB100-1771] - GFP antibody at a 1:1,000 dilution detects tau-GFP in cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) in Drosophila melanogaster late stage embryonic central nervous system. Fluorochrome conjugated anti-Goat secondary antibody was used for detection at 1:300. Panel A shows a lateral view (ventral left) and Panels B and C show ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). In all panels, anterior is up.
Western Blot: GFP Antibody [FITC] [NB100-1771] - Western blot of GFP recombinant protein detected with polyclonal anti-GFP antibody. Lane 1 shows detection of a 33 kDa band corresponding to a GFP containing recombinant protein (arrowhead) expressed in HeLa cells. Lane 2 shows no staining of a mock transfected HeLa cell lysate.
Western Blot: GFP antibody [NB600-303] - Recombinant GFP detected using NB 600-303. 10 second exposure.
Western Blot: GFP Antibody [NB600-308] - GFP protein detected with polyclonal anti-GFP antibody. Wild type GFP (0.1 ug) was used to spike 30 ug of HeLa whole cell lysate. Detects a 27 kDa band corresponding to the epitope tag GFP.
Immunocytochemistry/Immunofluorescence: GFP Antibody [NB600-310] - HEK 293 cells expressing GFP were visualized for GFP fluorescence (Green) or using NB600-310 to label the GFP (Red). Nuclei (Blue) were counterstained using Hoechst 33258.
Western Blot: GFP Antibody [NB600-310] - Detection of GFP in Western blot using NB 600-310. Lane 1: A549 + 500 ng GFP. Lane 2: A549 + 100 ng GFP. Lane 3: A549 + 20 ng GFP. Lane 4: 20 ng GFP. 10 ug of lysate were loaded and the primary dilution was 1:2,000. ECL detection was a 20 second exposure.
Western Blot: GFP Antibody (9F9.F9) [NB600-597] - Western blot of GFP recombinant protein detected with monoclonal anti-GFP antibody (NB600-597). GFP recombinant protein was expressed in HeLa cells, where 50 ng (Lanes 1), 100 ng (Lanes 2) and 500 ng (Lanes 3) of lysate were loaded per lane. NB600-597 detects a 27 kDa band corresponding to the epitope tag GFP. The cell lysates were prepared in a RIPA buffer containing 200 mM NaCl. The membrane was probed with the primary antibody diluted to 1.0 ug/ml for 1 h at room temperature followed by washes and reaction with a 1:2,500 dilution of IRDye800 conjugated Goat-a-Mouse IgG [H&L].
Western Blot: GFP Antibody (9F9.F9) [NB600-597] Known amounts of recombinant GFP protein were spiked into a HeLa cell-derived lysates and separated by SDS-PAGE using a 4-20% gradient gel. Proteins were transferred onto a nitrocellulose membrane for 1 h at 100 mV. The membrane was blocked with TTBS supplemented with 1% BSA (for 1 h at 4°C prior to probing the blot with the anti-GFP monoclonal antibody diluted 1:1,000 for 40 min. Detection of the primary antibody by the HRP-conjugated anti-Mouse IgG was performed at a dilution of 1:20,000 for 1h at 4°C. Super Sensitive Chemiluminescent Luminol Substrate was used for signal detection
Polyclonal anti-GFP antibody at a 1/1,000 dilution detects tau-GFP in cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) in Drosophila melanogaster late stage embryonic central nervous system. Fluorochrome conjugated anti-Goat secondary antibody was used for detection at 1/300. Panel A shows a lateral view (ventral left). Panels B and C show ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). In all panels, anterior is up.Personal Communication, Helmata Mistry, Washington University School of Medicine, St. Louis, MO.
Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus in Mus musculus (coronal view, 20X magnification). Briefly, Sf-1:Cre mice (Jackson Mouse Laboratories) were crossed to the Z/EG reporter line. Brains were harvested following cardiac perfusion with 4%PFA/PBS with o/n fixation, and subsequently transferred to a 30% sucrose solution. Brains were then frozen in OCT freezing medium (Sakura) and cryostat sectioned at 40 microns. Polyclonal Goat anti-GFP antibody was used at 1/500 dilution in free floating imunnohistochemistry to detect eGFP. Fluorchrome conjugated anti-goat IgG secondary antibody was used for detection at 1/500. Sections were counterstained with DAPI. Personal Communication, Daniel A. Lee, Johns Hopkins School of Medicine, Baltimore, MD.
Western blot of GFP recombinant protein detected with Polyclonal anti-GFP antibody. Lane 1 shows detection of a 33 kDa band corresponding to a GFP containing recombinant protein (arrowhead) expressed in HeLa cells. Lane 2 shows no staining of a mock transfected HeLa cell lysate. A 4-12% Bis-Tris gradient gel was used for SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking the membrane was probed with the primary antibody diluted to 1 µg/ml for 1 h at room temperature followed by washes and reaction with a 1/2,500 dilution of IRDye® 800 conjugated Donkey-a-Goat IgG [H&L]. The IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
Immunofluorescence Microscopy of Anti-GFP (GOAT) Antibody: Tissue: E5.5 Hex-GFP transgenic mouse embryo. Primary antibody: Goat anti-GFP was used at 1/500 dilution. Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody at 1/10,000 for 45 min at RT. Staining: GFP as green fluorescent signal with DAPI blue counterstain.
Immunofluorescence Microscopy of GFP-GOAT-Antibody. Tissue: Sf-1:Cre mice crossed to the Z/EG reporter line. Mouse brain (coronal view, 20X magnification). Fixation: 4%PFA/PBS with o/n fixation, and subsequently transferred to a 30% sucrose solution. Antigen retrieval: frozen in OCT freezing medium (Sakura) and cryostat sectioned at 40 microns. Primary antibody: Goat anti-GFP was used at 1/500 dilution in free floating imunnohistochemistry to detect GFP. Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody was used for detection at 1/500 at 1/10,000 for 45 min at RT. Localization: Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus. Staining: eGFP as green fluorescent signal and sections were counterstained with DAPI.
Immunofluorescence Microscopy: Polyclonal anti-GFP antibody at a 1/1,000 dilution detects tau-GFP in cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) in Drosophila melanogaster late stage embryonic central nervous system. AlexaFluor 488(TM) conjugated anti-Goat antibody was used for detection at 1/300. Panel A shows a lateral view (ventral left) and Panels B and C show ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). In all panels, anterior is up. Personal Communication, Helmata Mistry, Washington University School of Medicine, St. Louis, MO.
Western blot of GFP recombinant protein detected with Polyclonal anti-GFP antibody (R1091P). Lane 1 shows blot results where GFP recombinant protein was expressed in HeLa cells. Lane 2 shows control staining of HeLa lysate not expressing GFP. Polyclonal anti-GFP antibody detects a 33 kDa band corresponding to the epitope tag GFP. A 4-12% Bis-Tris gradient gel was used for SDS-PAGE. The protein was transferred to Nitrocellulose using standard methods. After blocking the membrane was probed with the primary antibody diluted to 1.0 µg/ml for 1 h at room temperature followed by washes and reaction with a 1/2,500 dilution of IRDye(R) 800 conjugated Donkey-a-Goat IgG [H&L]. IRDye(R)800 fluorescence image was captured using the Odyssey(R) Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
Western Blot of Anti-GFP (MOUSE) Monoclonal Antibody Peroxidase Conjugate. Lane 1: 50ng of GFP. Lane 2: none. Primary antibody: none. Secondary antibody: Anti-GFP (Mouse) Monoclonal Antibody Peroxidase Conjugate (Cat.-No R1461HRP) secondary antibody was used at 1:1000 for 45 min at RT. Block: 1% BSA-TTBS (p/n MB-013, diluted to 1X) 30 min at 20°C. Predicted/Observed size: 28 kDa for GFP. Other band(s): none.
Western blot of GFP recombinant protein detected with monoclonal anti-GFP antibody. GFP recombinant protein was expressed in HeLa cells, where 50 ng (lane 1), 100 ng (lane 2) and 500 ng (lane 3) of lysate were loaded per lane. Anti-GFP detects a 27 kDa band corresponding to the epitope tag GFP. The cell lysates were prepared in a RIPA buffer containing 200 mM NaCl. A 4-12% Bis-Tris gradient gel was used for SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking with 5% BLOTTO in PBS, the membrane was probed with the primary antibody diluted to 1.0 μg/ml for 1 h at room temperature followed by washes and reaction with a 1:2,500 dilution of IRDye® 800 conjugated Goat-a-Mouse IgG [H&L]IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
Lineage tracing of EGFP expressing migrating pectoral girdle myogenic precursors in the chicken embryos via electroporation using TOL-2-GFP plasmid system. Vibratome sections stained with anti-GFP antibody Cat.-No. TP401 show the labeled muscle fiber soft hepectoral girdle in chicken embryos at stage HH27.Data was kindly provided by: Nargis Khalida, Rizwan Rehimi: Institute of Anatomy, Dept. of Molecular Embryology, Ruhr University Bochum, Germany
Western blot detection of GFP-MIP2 fusion protein intransfectant by anti-GFP antibody.
Western blot analysis using GFP antibody. Approx MW ~26kDa
Left Panel - GFP immunostaining of neurons in the spinal cord of a transgenic mouse expressing GFP under the control of a gene product specifically expressed in laminae 2 of the dorsal horn gray matter. Right Panel - Western blot showing a single band at about 27 kDa, the molecular weight of GFP.
Left Panel - GFP immunostaining of neurons in the spinal cord of a transgenic mouse expressing GFP under the control of a gene product specifically expressed in laminae 2 of the dorsal horn gray matter. Right Panel - Western blot showing a single band at about 27 kDa, the molecular weight of GFP.
Western Blot of anti-GFP monoclonal antibody (GTX21218). Lane 1: 64pg of recombinant GFP protein were spiked into a HeLa cell-derived lysates). Lane 2: 32pg of recombinant GFP protein were spiked into a HeLa cell-derived lysates. Lane 3: 16pg of recombinant GFP protein were spiked into a HeLa cell-derived lysates. Lane 4: 8pg of recombinant GFP protein were spiked into a HeLa cell-derived lysates. Lane 5: 4pg of recombinant GFP protein were spiked into a HeLa cell-derived lysates. Lane 6: 2pg of recombinant GFP protein were spiked into a HeLa cell-derived lysates. Lane 7: 1g of recombinant GFP protein were spiked into a HeLa cell-derived lysates. Lane 8: 0pg of recombinant GFP protein were spiked into a HeLa cell-derived lysates. Primary antibody: anti-GFP monoclonal antibody at 1:400 for overnight at 4°C. Secondary antibody: HRP-conjugated anti-Mouse IgG (GTX26728) was performed at a dilution of 1:20,000 for 1h at 4°C. Block: TTBS supplemented with 1% BSA for 1 h at 4°C. Predicted/Observed size: 27 kDa for GFP. Other band(s): none.
Figure: Western blot of GFP recombinant protein detected with GeneTex monoclonal anti-GFP antibody. GFP recombinant protein was expressed in HeLa cells, where 50 ng (lane 1), 100 ng (lane 2) and 500 ng (lane 3) of lysate were loaded per lane. GTX21218 detects a 27 kDa band corresponding to the epitope tag GFP. The membrane was probed with the primary antibody diluted to 1.0 µg/ml for 1 h at room temperature followed by washes and reaction with a 1:2,500 dilution of IRDye® 800 conjugated goat anti-Mouse IgG [H&L]. IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
Immunofuorescence using anti-GFP antibody (GTX26556) to label GFP-expressing glial cells (green) transplanted into lesioned rat spinal cord. Axons are labelled red by an antibody to neurofilament-200 and a rhodamine secondary. Image courtesy of Andrew Toft.
Western Blot of Rabbit anti-GFP antibody (GTX26556). Lane 1: 293FT cells transfected with CDK4 dominant negative (C-). Lane 2: 293FT cells poitive control (C+). Load: 25 µg per lane. Primary antibody: GFP antibody at 1:400 for overnight at 4°C. Secondary antibody: Rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: 27 kDa for GFP.
Western blot of GFP protein detected with anti-GFP antibody (1:1000 dilution) and detected with IRDye® 800-conjugated goat anti-rabbit IgG (1:10,000 dilution). Wild type GFP (0.1 µg) was used to spike 30 µg of a HeLa whole cell lysate. This antibody detects a 27 kDa band corresponding to the epitope tag GFP.
Immuno-microscopy of Rabbit anti-GFP antibody (GTX26556). Monocyte derived dendritic cells and dermal macrophages were challenged and directly visualized with eGFP labeled Dengue virus to localize sequestration of virus particles in the different cells (upper). The location of the GFP was confirmed by TEM (lower magnified view) using GeneTex rabbit anti GFP Primary antibody (GTX26556) (1:200) and a gold labeled secondary antibody.
Western Blot of Rabbit anti-GFP antibody (GTX26658). Lane 1: HeLa cells. Lane 2: mock transfected HeLa cell lysate. Load: 35 µg per lane. Primary antibody: GFP antibody at 1 µg/ml for 1 h at room temperature. Secondary antibody: Infrared 800 conjugated Donkey-a-Goat IgG [H&L] MX7 secondary antibody at 1:2,500 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: 27 kDa, 33 kDa for GFP. Other band(s): none.
Western Blot of Alkaline Phosphatase conjugated Goat anti-GFP antibody (GTX26661). Lane 1: GFP. Lane 2: none. Load: 50 ng per lane. Primary antibody: none. Secondary antibody: Alkaline Phosphatase GFP secondary antibody at 1:1000 for 60 min at RT. Block for 30 min RT. Predicted/Observed size: 28 kDa for GFP. Other band(s): none.
Immunofluorescence Microscopy of GFP-GOAT-Antibody. Tissue: Sf-1:Cre mice crossed to the Z/EG reporter line. Mouse brain (coronal view, 20X magnification). Fixation: 4%PFA/PBS with o/n fixation, and subsequently transferred to a 30% sucrose solution. Antigen retrieval: frozen in OCT freezing medium (Sakura) and cryostat sectioned at 40 microns. Primary antibody: Goat anti-GFP (GTX26662) was used at 1:500 dilution in free floating imunnohistochemistry to detect GFP. Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody was used for detection at 1:500 at 1:10,000 for 45 min at RT. Localization: Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus. Staining: eGFP as green fluorescent signal and sections were counterstained with DAPI.
These pictures show confocal immunofluorescence using GFP-expressing glial cells (green) transplanted into the lesioned rat spinal cord. This was detected using GTX26662 and a standard FITC filter set. Axons are labelled red by an antibody to neurofilamen
Western Blot of Goat anti-GFP Fluorescein Conjugated Antibody (GTX26662).Lane 1: GFP. Lane 2: None. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Fluorescein goat secondary antibody at 1:1,000 for 60 min at RT. Block for 30 min at RT. Predicted/Observed size: 28 kDa, 33 kDa for GFP. Other band(s): None.
Western Blot of Anti-GFP Antibody Peroxidase Conjugate (GTX26663). Lane 1: 50ng of GFP. Lane 2: none. Primary antibody: none. Secondary antibody: Anti-GFP Antibody Peroxidase Conjugate secondary antibody was used at 1:1000 for 45 min at RT. Block: 1% BSA-TTBS 30 min at 20°C. Predicted/Observed size: 28 kDa for GFP. Other band(s): none
Figure 2. Immunofluorescence microscopy. GeneTex polyclonal anti-GFP antibody at a 1:1,000 dilution detects tau-GFP in cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) in Drosophila melanogaster late stage embryonic central nervous system. Fluorochrome conjugated anti-Goat secondary antibody was used for detection at 1:300. Panel A shows a lateral view (ventral left) and Panels B and C show ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). Personal Communication, Helmata Mistry, Washington University School of Medicine, St. Louis, MO.
Immunofluorescence Microscopy of Anti-GFP (GOAT) Antibody (GTX26673). Tissue: E5.5 Hex-GFP transgenic mouse embryo. Primary antibody: Goat anti-GFP was used at 1:500 dilution. Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody at 1:10,000 for 45 min at RT. Staining: GFP as green fluorescent signal with DAPI blue counterstain.
Figure 1. Western blot of GFP recombinant protein detected with GeneTex polyclonal anti-GFP antibody. Lane 1 shows detection of a 33 kDa band corresponding to a GFP containing recombinant protein (arrowhead) expressed in HeLa cells. Lane 2 shows no staining of a mock transfected HeLa cell lysate. After blocking the membrane was probed with the primary antibody diluted to 1 µg/ml for 1 h at room temperature followed by washes and reaction with a 1:2,500 dilution of donkey anti-Goat IgG [H&L].
Figure 3. Immunofluorescence microscopy. Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus in Mus musculus (coronal view, 20X magnification). GeneTex polyclonal goat anti-GFP was used at 1:500 dilution in free floating imunnohistochemistry to detect eGFP. Fluorchrome conjugated anti-goat IgG secondary antibody was used for detection at 1:500. Sections were counterstained with DAPI. Personal Communication, Daniel A. Lee, Johns Hopkins School of Medicine, Baltimore, MD.
Western blot detection of GFP-MIP2 fusion protein in transfectant by anti-GFP.
Western Blot: GFP Antibody (9F9.F9) [Biotin] [NB110-40670] detection of Green Fluorescent Protein. 50ng of GFP (Lane 1) was run on a 4-20% gel and transferred to 0.45 µm nitrocellulose. After blocking with 5% Blotto 30 min at 20°C, Anti-GFP (MOUSE) Monoclonal Antibody Biotin Conjugated secondary antibody was used at 1:5000 in Blocking Buffer for Fluorescent Western Blotting. HRP Streptavidin was used at 1:40,000 in buffer for 30 min at 20C
Immunoblot analysis of cell lysates. Lane 1 : Bosc23 cell lysate transfected with GFP-Ub, lane 2 : Bosc23 cell lysate transfected with GFP-Ub+1 and lane 3 : Bosc23 cell lysate transfected with GFP.
Confocal microscopy images of COS - 7 cells transfected with expression constructs encoding membrane - tethered EGFP ( membrane - EGFP ; top ) or nuclear Polycomb 2 - EYFP fusion protein ( Pc2 - EYFP ; bottom ) . The natural fluorescence of the produced proteins is shown in the green channel ( left ) , GFP polyclonal antibody ( Cat # PAB1264 ) signal was detected in the red channel ( right ) . The system was carefully tested for overlap of these two optical channels and images were scanned separately in sequential scanning mode. The blue nuclear stain is also shown.
Immunoprecipitation of GFP from HEK 293 cells using GFP polyclonal antibody ( Cat # PAB1264 ) . HEK 293 cells were transfected with expression construct encoding GFP protein. Twenty hours post transfection cells were lysed in non - denaturating conditions ( Lysis buffer : 20 mM Tris, pH 7.5, 100 mM NaCl, 0.5% Triton X - 100, inhibitors of proteases ) . Aliquots of cell lysate were immunoprecipitated using a GFP polyclonal antibody ( Cat # PAB1264 , lane 2 ) or a pre - immune rabbit serum ( lane 3 ) . Immunoprecipitates together with a sample of the cell lysate ( lane 1 ) were separated on SDS - PAGE polyacrylamide gel and immunoblotted with the anti - GFP antibody. The positions of molecular weight markers in kDa are indicated at the left.
Immunoprecipitation analysis of Bosc 23 cell lysates. Lane 1 : Input ( Bosc 23 cell lysate transfected with GFP ). Lane 2 : Precipitates. Immunoblotting with GFP tag monoclonal antibody, clone 18A11 ( Cat # MAB0849 ).
Formalin-fixed and paraffin-embedded mouse skin tissue expression GFP reacted with GFP Tag Antibody (Cat.-No AM11009PU-N ), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. -ve shows negative staining of the antibody in a wild type mouse, whereas +ve shows positive staining in a mouse expressing GFP under the LGR5 promoter. The staining shows localization of GFP at the bottom of the hair follicle. The antigen was retrieved in citrate pH 6 in water bath for 25 mins.
Immuno-fluorescence for GFP Tag Antibody (Cat.-No AM11009PU-N). Antibody target is a YFP tagged protein (the inner membrane complex) of Toxoplasma gondii, which localizes around the exterior of the parasite. Brighter parasites represent actively dividing cells and darker ones are not actively dividing. Secondary goat-anti-mouse and goat-anti-rabbit were conjugated with Alexa546 (Molecular Probes) at 1/200 dilution. Phase contrast enhancement applied. Fluorescent lamp exposure times as follows: RFP Channel, 0.075 sec; YFP Channel, 1.000 sec; DAPI Channel, 0.020 sec; Phase Channel: 0.300 sec. Microscope: Axiovert 200M inverted microscope (Zeiss) equipped with a PRIOR stage and a NEOFLUAR 100x/1.3 objective lens. Images were collected with a Hamamatsu C4742-95 CCD camera. Chroma Filtersets: TRITC, HQ545/30x Q570LP HQ620/60m; DAPI, D350/50x E420LP 400DCLP; YFP, Q151LP HQ500/20x HQ535/30m. Data courtesy of Seth Robertson, Boston College.
Top: Western blot analysis of Monoclonal anti-GFP antibody (Cat.-No AM11009PU-N) using purified GFP, YFP and BFP proteins expressed in bacteria: Both GFP (Lanes 1-3) and YFP (Lanes 4-6) but not BFP (data not shown) were detected using the purified Mab. Buttom: Western blot analysis of anti-GFP Tag Antibody (SG071130A) in GFP recombinant protein. GFP recombinant protein (arrow) was detected using the purified Mab.
Western blot analysis of anti-GFP Tag Antibody Cat.-No AM11010SU-N (Clone CA071114E) using purified GFP, YFP and BFP proteins expressed in bacteria: Both GFP (Lanes 1-3) and YFP (Lanes 4-6) but not BFP (data not shown) were detected using the purified Mab.
Western blot analysis of anti-GFP Tag Antibody Cat.-No AM11010SU-N (Clone CA071114E) in GFP recombinant protein. GFP recombinant protein (arrow) was detected using the purified Mab.
Analysis:1.Prepare a 150 ul recombinant GFP with various concentrations (0.00074 nM - 6.21 uM) in assay buffer and equilibrate to 25°C. (Assay buffer: 10 mM Tris-HCl (pH 8.0), 10 mM EDTA, 0.02% sodium azide.)2.Read at excitation wavelengths 485nm and emission 535nm. - 96 Well Polystyrene Microplate, black (greiner bio-one, Cat.No, 655077)- Fluorescent plate reader (PerkinEmerl, VICTOR X3)
GFP, 1-238 aa Aequorea victoria, Recombinant, E.coli: 15% SDS-PAGE (3 µg)
Western blot: The recombinant protein GFP (20, 40 and 80 ng) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-GFP antibody (1/1000). Proteins were visualized using a goat anti-Mouse secondary antibody conjugated to HRP and an ECL detection system. Lane 1: GFP 20 ng Lane 2: GFP 40 ng Lane 3: GFP 80 ng
Western blot analysis of GFP expression in 293 cells transfected with a GFP-fusion construct.
Western blot of GFP recombinant protein detected with GFP monoclonal antibody ( Cat # MAB2003 ).GFP recombinant protein was expressed in HeLa cells, where 50 ng ( lane 1 ), 100 ng ( lane 2 ) and 500 ng ( lane 3 ) of lysate were loaded per lane.GFP monoclonal antibody ( Cat # MAB2003 ) detects a 27 kDa band corresponding to the epitope tag GFP.The cell lysates were prepared in a RIPA buffer containing 200 mM NaCl.A 4-12% Bis-Tris gradient gel ( Invitrogen ) was used for SDS-PAGE.The protein was transferred to nitrocellulose using standard methods.After blocking with 5% BLOTTO in PBS, the membrane was probed with the primary antibody diluted to 1.0 µg/ml for 1 h at room temperature followed by washes and reaction with a 1 : 2,500 dilution of IRDye® 800 conjugated Goat-a-Mouse IgG [H&L] MX10.IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR.IRDye is a trademark of LI-COR, Inc.
Immunofluorescence microscopy of GFP polyclonal antibody ( Cat # PAB10341 ) at a 1 : 1,000 dilution detects tau-GFP in cell bodies ( large arrowhead ) and axons of motorneurons ( arrow ) and interneurons ( small arrowhead ) in Drosophila melanogaster late stage embryonic central nervous system.AlexaFluor 488 conjugated anti-Goat antibody was used for detection at 1 : 300.Panel A shows a lateral view ( ventral left ) and Panels B and C show ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom).Personal Communication, Helmata Mistry, Washington University School of Medicine, St. Louis, MO.
Western blot of GFP recombinant protein detected with GFP polyclonal antibody ( Cat # PAB10341 ).Lane 1 shows detection of a 33 kDa band corresponding to a GFP containing recombinant protein ( arrowhead ) expressed in HeLa cells.Lane 2 shows no staining of a mock transfected HeLa cell lysate.A 4-12% Bis-Tris gradient gel was used for SDS-PAGE.The protein was transferred to nitrocellulose using standard methods.After blocking the membrane was probed with the primary antibody diluted to 1.0 µg/ml for 1 h at room temperature followed by washes and reaction with a 1 : 2,500 dilution of IRDye® 800 conjugated Donkey-a-Goat IgG [H&L] MX7.The IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR.IRDye is a trademark of LI-COR, Inc.
Western blot of GFP protein detected with GFP polyclonal antibody ( Cat # PAB10342 ).Wild type GFP ( 0.1 µg ) was used to spike 30 µg of a HeLa whole cell lysate.This antibody detects a 27 kDa band corresponding to the epitope tag GFP.A 4-20% Tris-Glycine gradient gel was used for SDS-PAGE.The protein was transferred to nitrocellulose using standard methods.After blocking with 5% BLOTTO in PBS, the membrane was probed overnight at 4°C with the primary antibody diluted in 5% BLOTTO to 1 : 1,000, followed by washes and reaction with a 1 : 10,000 dilution of IRDye® 800 conjugated Goat-a-Rabbit IgG [H&L] MX10.IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR.IRDye is a trademark of LI-COR, Inc.
The cell lysate of 293T with over-expressed recombinant protein GFP was resolved onto 10% SDS-PAGE, then transferred onto NC membrane. Followed by an immune-blotting with GFP polyclonal antibody ( Cat # PAB12768 ) at 1 : 1000.
Western blot analysis of EGFP polyclonal antibody ( Cat # PAB8931 ) using recombinant RFP ( dsRed, Lane 1 and 2 ), YFP ( Lane 3 and 4 ) and EGFP ( Lane 5 and 6 ).
Immunofluorescence analysis of GFP-transfected HeLa. A:GFP is expressed in the tranfected cell. B: The cell expressing GFP can be detected using rabbit anti-GFP ab ( GTX113617, 1:5000) followed by Alexa Fluor 594 (1:500) goat anti-rabbit IgG. C: Merged with DNA probe, the lower cell with no GFP expressed as a negtive control.
GFP antibody detects GFP protein by Western blot analysis.A. 1 µg 293T whole cell extract B. 1 µg 293T whole cell extract expressing GFP-tagged protein C. 1 µg 293T whole cell extract expressing RFP-tagged protein 12 % SDS-PAGEGFP antibody (GTX113617) dilution: 1:10000
IP-WB assay to show that Hice1 (GTX113617) co-immunoprecipitated with other Augmin components HAUS6 (GTX118732), HAUS2 (GTX118734) and HAUS1 (GTX118733) in U2OS cells.
GFP was immunoprecipitated from bacterial GFP-expressing lysate using 1 ug of anti-GFP antibody (GTX113617). The precipitated GFP was detected by GTX113617 diluted at 1:15000.
Immunohistochemical analysis (whole mount) of transgenic zebrafish embryo, using GFP(GTX113617) antibody at 1:200 dilution.
Western blot of GFP protein detected with polyclonal anti-GFP antibody. Wild type GFP (0.1 μg) was used to spike 30 μg of a HeLa whole cell lysate. This antibody detects a 27 kDa band corresponding to the epitope tag GFP. A 4-20% Tris-Glycine gradient gel was used for SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking with 5% BLOTTO in PBS, the membrane was probed overnight at 4° C with the primary antibody diluted in 5% BLOTTO to 1:1,000, followed by washes and reaction with a 1:10,000 dilution of IRDye® 800 conjugated Goat-a-Rabbit IgG [H&L]. IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
Western blot analysis of recombinant protein GFP ( 20, 40 and 80 ng ) were resolved by SDS-PAGE, transferred to NC membrane and probed with GFP monoclonal antibody, clone AT2G5 ( Cat # MAB5377 ) ( 1 : 1000 ).Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.Lane 1 : GFP 20 ng.Lane 2 : GFP 40 ng.Lane 3 : GFP 80 ng.
Western Blot of Rabbit anti-GFP antibody: Lane 1: Wild type GFP (0.1 µg) was used to spike HeLa whole cell lysate. Lane 2: none. Load: 30 µg per lane. Primary antibody: GFP antibody at 1:1000 for overnight at 4°C. Secondary antibody: IRDye800™ Goat-a-Rabbit IgG [H&L] MX10 at 1:10,000 for 45 min at RT. Block: 5% BLOTTO in PBS overnight at 4°C.
Predicted/Observed size: 27 kDa for epitope tag GFP. Other band(s): none.
Predicted/Observed size: 27 kDa for epitope tag GFP. Other band(s): none.
Western blot of GFP protein detected with polyclonal anti-GFP antibody. Lane 1 shows negative control staining of 20 µg of mouse spleen lysate. Lane 2 shows staining of mouse spleen lysate spiked with 50 ng of wt GFP. This antibody detects a 27 kDa band corresponding to the GFP epitope tag commonly used in recombinant constructs. A 4-20% Tris-Glycine gradient gel was used for SDS-PAGE followed by transfer to nitrocellulose using standard methods. After blocking with 5% BSA in PBS, the membrane was probed for 2 h at room temperature with the primary antibody diluted in 5% BSA to 2 µg/ml, followed by washes and reaction with a 1:20,000 dilution of IRDye(TM)800 Conjugated AffinityPurified anti-Chicken IgG [H&L] [Goat] MX10. The IRDye®(R)800 fluorescence image was captured using the Odyssey(R) Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
Western blot analysis of over-expressed GFP-tagged protein in 293T cell lysate, using GFP-Tag (7G9) Monoclonal Antibody.
IP of extracts from 293T cells transfected (lane 1,2) with a GFP-tagged protein and a mock transfection (lane 3).
Immunofluorescence analysis of 293T cells transfected with a GFP-tagged protein, using GFP-Tag (7G9) Mouse mAb at a 1:500 dilution.
Western blot analysis of over-expressed GFP-tagged protein in 293T cell lysate, using GFP-tag (7G9) Mouse mAb. The antibody dilutions are 1:5000 (lane 1) and 1:10000 (lane 2). Each lane was loaded with 10 μg of cell lysate.
Immunoprecipitation of extracts from 293T cells transfected (lane 1,2) with a GFP-tagged protein and a mock transfection (lane 3, using the same protein without the GFP tag), using GFP-Tag (7G9) Mouse mAb and probed on Western blot using the same antibody. Dilution: 1:50 (1), 1:100 (2), 1:50 (3).
Antibody working Cont: 1/1000 from 1 mg/ ml stock Cont.
Lane 1: Negative Control. Lane 2: 1 μg GFP Fusion Protein Lysate Lane 3: 10 μg GFP Fusion Protein Lysate
Antibody working Cont: 1/1000 from 1 mg/ ml stock Cont.
Lane 1: Negative Control. Lane 2: 1 μg GFP Fusion Protein Lysate. Lane 3: 10 μg GFP Fusion Protein Lysate.
GFP antibody (HRP) detects GFP protein by Western blot analysis.A. 4 µg HeLa whole cell extractB. 4 µg HeLa whole cell extract expressing GFP-tagged protein 12 % SDS-PAGEGFP antibody (HRP) (GTX113618-01) dilution: 1:4000
The same murine cortical neuron visualized using our Chicken anti-GFP antibody (left, in red) and visualized using GFP autofluorescence (right, in green). Note the increased detail and intensity of the anti-GFP antibody staining.
Immunocytochemistry analysis of HCC827 cells, untransfected (left) or transfected with GFP (right) using GFP antibody Cat.-No AM06180PU-N
Western blot analysis using GFP antibody Cat.-No AM06180PU-N against extracts from HCC827 cells, untransfected (Lane 1) and transfected with GFP (Lane 2).
Western blot analysis using GFP antibody Cat.-No AM06180PU-N against recombinant GFP fusion protein (Lane 1) and various recombinant fusion protein with GFP tag (Lanes 2, 3, 4).
Flow Cytometric analysis of HCC827 cells, untransfected (blue) or transfected with GFP (green), using GFP GFP antibody Cat.-No AM06180PU-N
Immunohistochemical detection of GFP on paraffin embedded section of GFP transgenic mouse with AM26599AF-N.
Immunohistochemical detection of GFP on paraffin embedded section of GFP transgenic mouse.
Magenta: AM26599AF-N.
Green: GFP own fluorescence.
Cyan, Blue: DAP.
Magenta: AM26599AF-N.
Green: GFP own fluorescence.
Cyan, Blue: DAP.
Immunocytochemical detection of GFP on 4% PFA fixed transfectant using AM26599AF-N (left). Right panel is GFP own fluorescence.
Immunoprecipitation of GFP from transfectant
Lane 1: IP with AM26599AF-N.
Lane 2: IP with isotype control.
Lane 1: IP with AM26599AF-N.
Lane 2: IP with isotype control.
Recombinant protein were subjected to SDS PAGE followed by western blot with HRP-66002(GFP tag Antibody) at dilution of 1:10000
Formalin-fixed and paraffin embedded pig liver labeled with Anti-GFP Polyclonal Antibody, FITC Conjugated (bs-0890R-FITC) at 1:200 for 60 min at 37C, followed by labeling with Anti-CK18 Polyclonal Antibody, Cy5 Conjugated (bs-2043R-Cy5) at 1:200 for 60 min at 37C and DAPI staining
Western analysis of recombinant GFP-His protein using a rabbit polyclonal anti-GFP antibody CAt.-No AP33450PU-N
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Primary Antibodies
| Catalog No. | Host | Iso. | Clone | Pres. | React. | Applications | |
|---|---|---|---|---|---|---|---|
| SP3005P | GFP antibody |
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Rabbit | Purified | All Species | ICC/IF, IP, WB |
0.1 mg /
€230.00
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| OriGene Technologies GmbH | |||||||
 +1 additional image |
Top Selling product! Published recently! |
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| AM11009PU-N | GFP antibody |
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Mouse | IgG1 | 168AT1211 | Purified | ICC/IF, P, WB |
0.4 ml /
€415.00
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| OriGene Technologies GmbH | |||||||
| +2 additional images |
Published recently! |
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| AM11010SU-N | GFP antibody |
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Mouse | IgG | 168AT1211.269.64 | Ascites | WB |
0.1 ml /
€415.00
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| OriGene Technologies GmbH | |||||||
| +1 additional image |
Published recently! |
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| AP09218PU-N | GFP (Ads. to Hu, Ms, Rt Serum Proteins) antibody |
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![Western blot of GFP protein detected with polyclonal anti-GFP antibody. Wild type GFP (0.1 μg) was used to spike 30 μg of a HeLa whole cell lysate. This antibody detects a 27 kDa band corresponding to the epitope tag GFP. A 4-20% Tris-Glycine gradient gel was used for SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking with 5% BLOTTO in PBS, the membrane was probed overnight at 4° C with the primary antibody diluted in 5% BLOTTO to 1:1,000, followed by washes and reaction with a 1:10,000 dilution of IRDye® 800 conjugated Goat-a-Rabbit IgG [H&L]. IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.](https://m1.acris-antibodies.com/catalog/product/cache/1/small_image/50x50/9df78eab33525d08d6e5fb8d27136e95/0163839.png) |
Rabbit | IgG | Aff - Purified | A. victoria | C, E, ICC/IF, WB |
0.1 mg /
€390.00
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| OriGene Technologies GmbH | |||||||
| +2 additional images |
Published recently! |
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| AP54983SU-N | GFP antibody |
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Rabbit | Serum | C, E, ICC/IF, WB |
0.2 ml /
€380.00
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| OriGene Technologies GmbH | |||||||
Published recently! |
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| R1091B | GFP (Ads. to Hu, Ms, Rt Serum Proteins) antibody |
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Goat | Biotin | A. victoria | C, E, ICC/IF, WB |
1 mg /
€630.00
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| OriGene Technologies GmbH | |||||||
| +2 additional images |
Published recently! |
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| R1091F | GFP antibody |
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Goat | FITC | A. victoria | C, E, ICC/IF, WB |
1 mg /
€640.00
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| OriGene Technologies GmbH | |||||||
Published recently! |
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| R1091P | GFP (Ads. to Hu, Ms, Rt Serum Proteins) antibody |
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Goat | Aff - Purified | C, E, ICC/IF, WB |
1 mg /
€600.00
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| OriGene Technologies GmbH | |||||||
| +3 additional images |
Published recently! Highly validated antibody! |
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| R1091T | GFP antibody |
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Goat | TRITC | E, ICC/IF, WB |
1 mg /
€610.00
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| OriGene Technologies GmbH | |||||||
Published recently! |
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| R1091TR | GFP antibody |
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Goat | Texas Red | C, ICC/IF |
1 mg /
€630.00
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| OriGene Technologies GmbH | |||||||
Published recently! |
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| R1461HRP | GFP antibody |
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Mouse | IgG2a | 9F9.F9 | HRP | A. victoria | C, E, WB |
1 mg /
€855.00
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| OriGene Technologies GmbH | |||||||
Published recently! |
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| AM06180PU-N | GFP antibody |
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Mouse | IgG2a | 4B10B2 | Purified | E, F, P, WB |
0.1 ml /
€340.00
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| OriGene Technologies GmbH | |||||||
| +3 additional images |
Highly validated antibody! |
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| AM26599AF-N | GFP antibody |
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Mouse | IgG2b | 1E4 | Azide Free | ICC/IF, IP, P, WB |
0.1 mg /
€355.00
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| OriGene Technologies GmbH | |||||||
| +4 additional images |
Highly validated antibody! |
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| 73-131 | GFP antibody |
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Mouse | IgG2a | N86/8 | Supernatant | C, ICC/IF, IP, WB |
5 ml /
€490.00
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| Antibodies Incorporated | |||||||
| +2 additional images |
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| 73-132 | GFP antibody |
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Mouse | IgG2a | N86/38 | Supernatant | C, ICC/IF, IP, WB |
5 ml /
€490.00
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| Antibodies Incorporated | |||||||
| +2 additional images |
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| 75-131 | GFP antibody |
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Mouse | IgG2a | N86/8 | Purified | C, ICC/IF, IP, WB |
0.1 ml /
€570.00
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| Antibodies Incorporated | |||||||
| +2 additional images |
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| 75-132 | GFP antibody |
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 |
Mouse | IgG2a | N86/38 | Purified | C, ICC/IF, IP, WB |
0.1 ml /
€570.00
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| Antibodies Incorporated | |||||||
| +2 additional images |
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| AB0020-20 | GFP antibody |
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Goat | IgG | Aff - Purified | C, ICC/IF, P, WB |
20 µl /
€50.00
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| SICGEN | |||||||
| AB0020-200 | GFP antibody |
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Goat | IgG | Aff - Purified | C, ICC/IF, P, WB |
0.4 mg /
€210.00
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| SICGEN | |||||||
| AB0020-500 | GFP antibody |
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Goat | IgG | Aff - Purified | C, ICC/IF, P, WB |
1.0 mg /
€420.00
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| SICGEN | |||||||
