AOF2 antibody
Principal name
AOF2 antibody
Alternative names for AOF2 antibody
Lysine-specific histone demethylase 1, KIAA0601, LSD1, Flavin-containing amine oxidase domain-containing protein 2, BRAF35-HDAC complex protein BHC110
Ncbi ID
9606, NP_001009999.1, NP_055828, NM_015013, NM_001009999, NP_001009999, NP_598633.2, NM_133872
Available reactivities
Available hosts
Available applications
Enzyme Immunoassay (E), Western blot / Immunoblot (WB), Paraffin Sections (P), ELISA (detection) (E(detection)), Immunocytochemistry/Immunofluorescence (ICC/IF), Frozen Sections (C), ChIP assay (ChIP), Immunoprecipitation (IP)
Background of AOF2 antibody
Lysine-specific demethylase 1 (LSD1; also known as AOF2 and BHC110) is a nuclear amine oxidase homolog that acts as a histone demethylase and transcription cofactor. Gene activation and repression is specifically regulated by the methylation state of distinct histone protein lysine residues. For example, methylation of histone H3 at Lys4 facilitates transcriptiol activation by coorditing the recruitment of BPTF, a component of the NURF chromatin remodeling complex, and WDR5, a component of multiple histone methyltransferase complexes. In contrast, methylation of histone H3 at Lys9 facilitates transcriptiol repression by recruiting HP1. LSD1 is a component of the CoREST transcriptiol co-repressor complex that also contains CoREST, CtBP, HDAC1 and HDAC2. As part of this complex, LSD1 demethylates mono-methyl and di-methyl histone H3 at Lys4 through a FAD-dependent oxidation reaction to facilitate neurol-specific gene repression in non-neurol cells. In contrast, LSD1 associates with androgen receptor in human prostate cells to demethylate mono-methyl and di-methyl histone H3 at Lys9 and facilitate androgen receptor-dependent transcriptiol activation. Therefore, depending on gene context LSD1 can function as either a transcriptiol co-repressor or co-activator. LSD1 activity is inhibited by the amine oxidase inhibitors pargyline, deprenyl, clorgyline and tranylcypromine.
Immunoperoxidase of monoclonal antibody to AOF2 on formalin-fixed paraffin-embedded human kidney. [antibody concentration 1 ug/ml]
Detection limit for recombinant GST tagged AOF2 is approximately 0.3ng/ml as a capture antibody.
Formalin-fixed and paraffin-embedded human breast cancer tissue reacted with AOF2 polyclonal antibody ( Cat # PAB1843 ) , which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
Western blot analysis of AOF2 ( arrow ) using AOF2 polyclonal antibody ( Cat # PAB1843 ) . 293 cell lysates ( 2 µg/lane ) either nontransfected ( Lane 1 ) or transiently transfected with the AOF2 gene ( Lane 2 ) ( Origene Technologies ) .
Western blot analysis of AOF2 polyclonal antibody ( Cat # PAB1843 ) in mouse brain tissue lysate ( 35 µg/lane ) . AOF2 ( arrow ) was detected using the purified polyclonal antibody.
Formalin-fixed and paraffin-embedded human hepatocellular cancer tissue reacted with AOF2 polyclonal antibody ( Cat # PAB1844 ) , which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
Western blot analysis of AOF2 ( arrow ) using AOF2 polyclonal antibody ( Cat # PAB1844 ) . 293 cell lysates ( 2 µg/lane ) either nontransfected ( Lane 1 ) or transiently transfected with the AOF2 gene ( Lane 2 ) ( Origene Technologies ) .
Formalin-fixed and paraffin-embedded human hepatocellular cancer tissue reacted with AOF2 polyclonal antibody ( Cat # PAB1845 ) , which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
Western blot analysis of AOF2 ( arrow ) using AOF2 polyclonal antibody ( Cat # PAB1845 ) . 293 cell lysates ( 2 µg/lane ) either nontransfected ( Lane 1 ) or transiently transfected with the AOF2 gene ( Lane 2 ) ( Origene Technologies ) .
Immunofluorescence of purified MaxPab antibody to AOF2 on HeLa cell. [antibody concentration 10 ug/ml]
AOF2 MaxPab polyclonal antibody. Western Blot analysis of AOF2 expression in human pancreas.
Western Blot analysis of AOF2 expression in transfected 293T cell line (H00023028-T01) by AOF2 MaxPab polyclonal antibody.Lane 1: AOF2 transfected lysate(96.36 KDa).Lane 2: Non-transfected lysate.
Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
Western blot analysis of AOF2 (arrow) using LSD1 Antibody (Center) (Cat.#AP11212PU-N). 293 cell lysates (2 ug/lane) either nontransfected (Lane 1) or transiently transfected with the AOF2 gene (Lane 2) (Origene Technologies).
Brain, cortex: Formalin-Fixed Paraffin-Embedded (FFPE)
Heart: Formalin-Fixed Paraffin-Embedded (FFPE)
Western blot analysis of AOF2 in P815 cell lysate with AOF2 polyclonal antibody ( Cat # PAB13189 ) at ( A ) 1 and ( B ) 2 µg/ml.
Immunohistochemistry of AOF2 in human small intestine tissue with AOF2 polyclonal antibody ( Cat # PAB13190 ) at 2 µg/ml.
Western blot analysis of AOF2 in P815 cell lysate with AOF2 polyclonal antibody ( Cat # PAB13190 ) at ( A ) 1 and ( B ) 2 µg/ml.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma and kidney carcinoma tissue, showing nuclear localization using AOF2 monoclonal antibody, clone 1B2E5 ( Cat # MAB5530 ) with DAB staining.
Western blot analysis using AOF2 monoclonal antibody, clone 1B2E5 ( Cat # MAB5530 ) against truncated AOF2 recombinant protein ( Lane 1 ), HeLa ( Lane 2 ) and Jurkat ( Lane 3 ) cell lysate.
Western blot was performed using AOF2 polyclonal antibody ( Cat # PAB14107 ) diluted 1 : 1,000 in TBS-Tween containing 5% skimmed milk.The antibody recognizes endogenous AOF2 in HeLa NE ( 20 µg ).
Western blot was performed using AOF2 polyclonal antibody ( Cat # PAB14107 ) diluted 1 : 1,000 in TBS-Tween containing 5% skimmed milk. The antibody recognizes the recombinant AOF2 ( 50 ng ) ( Lane 1 ) as well as endogenous AOF2 in nuclear extracts from HeLa ( Lane 3 ) and HEK293 ( Lane 4 ).
Western blot was performed using nuclear extracts from HeLa cells ( HeLa NE, 40 µg ) and AOF2 polyclonal antibody ( Cat # PAB14121 ) diluted 1 : 1,000 in TBS-Tween containing 5% skimmed milk.
ELISA was performed using a serial dilution of AOF2 polyclonal antibody ( Cat # PAB14121 ), crude serum and Flow Through in antigen coated wells.By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1 : 20,000.
293T cells were transiently transfected with an expression vector for HA-tagged AOF2 ( Lanes 1 ) or with an empty vector ( Lanes 2 ).A : Whole cell extracts were analysed by Western blot using AOF2 polyclonal antibody ( Cat # PAB14134 ), diluted 1 : 1,000.B. Whole cell extracts were immunoprecipitated with 2 µg of AOF2 polyclonal antibody ( Cat # PAB14134 ). Immunoprecipitates were analysed by Western blot as described above.
Western blot was performed using respectively 150 µg ( Lane 1 ) or 50 µg ( Lane 2 ) nuclear extracts from HeLa cells and AOF2 polyclonal antibody ( Cat # PAB14134 ) diluted 1 : 1,000.
ELISA was performed using a serial dilution of AOF2 polyclonal antibody ( Cat # PAB14134 ).The plates were coated with the peptide used for immunization of the rabbit.By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1 : 2,500.
Immunoprecipitation of AOF2 transfected lysate using anti-AOF2 MaxPab rabbit polyclonal antibody and Protein A Magnetic Bead (U0007), and immunoblotted with AOF2 MaxPab mouse polyclonal antibody (B01) (H00023028-B01).
Human Skin: Formalin-Fixed, Paraffin-Embedded (FFPE)
Western blot analysis of in P815 cell lysate with KDM1A (LSD1) antibody (AP22944PU-N) at (A) 1 and (B) 2 µg/ml.
Human Skin: Formalin-Fixed, Paraffin-Embedded (FFPE)
Human Tonsil: Formalin-Fixed, Paraffin-Embedded (FFPE)
Western blot analysis in P815 cell lysate with KDM1A (LSD1) antibody (AP22945PU-N) at (A) 1 and (B) 2 µg/ml.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma (left) and kidney carcinoma (right), showing nuclear localization using LSD1 mouse mAb with DAB staining.
Western blot analysis using LSD1 mouse mAb against COS (1), Hela (2), NIH/3T3 (3), A549 (4) and Jurkat (5) cell lysate.
Western blot analysis of LSD1 in P815 cell lysate with LSD1 antibody at (A) 1 and (B) 2 µg/ml.
Western blot analysis of LSD1 in P815 cell lysate with LSD1 antibody at (A) 1 and (B) 2 ug/ml.
Immunohistochemistry of LSD1 in human small intestine tissue with LSD1 antibody at 2 ug/ml.
Immunofluorescence of LSD1 in Human Small Intestine cells with LSD1 antibody at 20 ug/mL.
Western blot analysis of LSD1/AOF2 using Jurkat cell lysate
Predicted band size: 95 kDa . Positive control: SW480 and PC-3 cell lysate. Recommended dilution: 1/500-2000. (Gel: 8%SDS-PAGE Lane1: SW480 cell lysate Lane2: PC-3 cell lysate Lysates: 40 ug per lane Primary antibody: 1/500 dilution Secondary antibody: Goat anti Rabbit IgG - H&L (HRP) at 1/10000 dilution Exposure time: 1 minute)
Western blot analysis of anti-LSD1 Pab (Cat. #TA324483) in mouse brain tissue lysate (35ug/lane). LSD1(arrow) was detected using the purified Pab.
Western blot analysis of AOF2 (arrow) using LSD1 Antibody (N-term) (Cat.#TA324483).293 cell lysates (2 ug/lane) either nontransfected (Lane 1) or transiently transfected with the AOF2 gene (Lane 2) (Origene Technologies).
Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.
IF image of Hela cell stained with LSD1 Antibody (N-term)(Cat#TA324483).Hela cells were incubated with LSD1 primary antibody (1:25, 1 h at 37?). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:400).Cytoplasmic actin was counterstained with Alexa Fluor® 555 (red) conjugated Phalloidin (7 units/ml). Nuclei were counterstained with DAPI (blue) . LSD1 immunoreactivity is localized to Cytoplasm significantly and Nucleus weakly.
Western blot analysis of AOF2 (arrow) using LSD1 Antibody (C-term) (Cat.#TA324484). 293 cell lysates (2 ug/lane) either nontransfected (Lane 1) or transiently transfected with the AOF2 gene (Lane 2) (Origene Technologies).
Western immunoblot. Nuclear extracts of control and PhIP-treated HMEC. Proteins were transferred onto polyvinylidene difluoride and blotted with anti-LSD1 antibody. Nuclear LSD1 protein levels increased in carcinogen-treated HMEC compared with control HMEC.
Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.
IF image of Hela cell stained with hLSD1-Y712(Cat#TA324484). Hela cells were incubated with hLSD1 primary antibody (1:25, 1 h at 37?). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:400).Cytoplasmic actin was counterstained with Alexa Fluor® 555 (red) conjugated Phalloidin (7 units/ml). Nuclei were counterstained with DAPI (blue) . hLSD1 immunoreactivity is localized to nucleus significantly.
Western blot analysis of extracts of PC12 cell lysate using KDM1A antibody.
Immunohistochemistry of paraffin-embedded human liver cancer using KDM1A antibody at dilution of 1:100 (400x lens).
Immunohistochemistry of paraffin-embedded mouse kidney using KDM1A antibody at dilution of 1:100 (400x lens).
Immunohistochemistry of paraffin-embedded rat brain using KDM1A antibody at dilution of 1:100 (400x lens).
Immunohistochemistry of paraffin-embedded rat kidney using KDM1A antibody at dilution of 1:100 (400x lens). .
Immunofluorescence analysis of HeLa cell using KDM1A antibody. Blue: DAPI for nuclear staining
TA329082 (0.5µg/ml) staining of Human Kidney lysate (35µg protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence.
Western Blot: LSD1 Antibody (1B2E5) [TA336270] - Analysis of LSD1 expression in Hela (1) and Jurkat (2) whole cell lysates.
Western Blot: LSD1 Antibody (1B2E5) [TA336270] - Western blot analysis using LSD1 mouse mAb against COS (1), Hela (2), NIH/3T3 (3), A549 (4) and Jurkat (5) cell lysate.
Immunohistochemistry-Paraffin: LSD1 Antibody (1B2E5) [TA336270] - Immunohistochemical analysis of paraffin-embedded Human Kidney Carcinoma tissue, showing nuclear localization using LSD1 antibody with DAB staining.
Immunohistochemistry-Paraffin: LSD1 Antibody (1B2E5) [TA336270] - Immunohistochemical analysis of paraffin-embedded Human Lung Carcinoma tissue, showing nuclear localization using LSD1 antibody with DAB staining.
Western blot detection of KDM1/LSD1 in Hela,HCT116 and COS7 cell lysates using KDM1/LSD1 mouse mAb (1:1000 diluted).Predicted band size:110KDa.Observed band size:110KDa.
Immunocytochemistry staining of HeLa cells fixed with 4% Paraformaldehyde and using anti-KDM1/LSD1 mouse mAb (dilution 1:100).
Immunoprecipitation analysis of Hela cell lysates using KDM1/LSD1 mouse mAb.
WB was performed on nuclear extracts from HeLa cells (20 ug) using the antibody against LSD1 diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (in kDa) is shown on the left; the location of the protein of interest is indicated on the right.
WB was performed using nuclear extracts from HeLa cells (40 ug) and the antibody against LSD1 diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (in kDa) is shown on the left. The location of the protein of interest is indicated on the right.
ChIP was performed with the ab against LSD1 on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 ug per ChIP experiment was analysed. IgG (2 ug/IP) was used as negative control. qPCR primers were for specific regions in the MYT1, RBM19, and TGFBR3 genesas positive controls, and for the MYOD1 gene, used as negative control. Image shows the recovery, expressed as a % of input (the relative amount of IP'd DNA compared to input DNA after qPCR analysis).
ChIP was performed on sheared chromatin from 4,000,000 K562 cells using 5 ug ab. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. The 50 bp tags were aligned to the human genome using the BWA algorithm. Image shows the peak distribution along the complete sequence and a 3 Mb region of the X-chromosome and in three regions surrounding the MYT1, RBM19 and TGFBR3 positive controls, respectively (C, D and E). The position of the amplicon used for ChIP-qPCR is indicated by arrow.
Determination of the antibody titer To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against LSD1 in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:20,000.
HeLa cells were stained with the antibody against LSD1 and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the LSD1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
WB was performed using respectively 150 ug (lane 1) or 50 ug (lane 2) nuclear extracts from HeLa cells and the antibody against LSD1 diluted 1:1,000. The location of the protein of interest is indicated on the left.
Determination of the antibody titer To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against human LSD1. The plates were coated with the peptide used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:2,500.
Immunoprecipitation and WB using the antibody against human LSD1 (cat. No. pAb-125-050), diluted 1:1,000. Figure 3B. Whole cell extracts were immunoprecipitated with 2 ug of the antibody against LSD1. Immunoprecipitates were analysed by WB as described above.
WB was performed with the antibody against LSD1 diluted 1:1,000 in TBS-Tween containing 5% skimmed milk on purified recombinant LSD1 protein (50 and 25 ng, lane 1 and 2) and on nuclear extracts (50 ug) from HeLa and HEK293 cells (lane 3 and 4, respectively). Figure 1B: WB was performed as described above using nuclear extracts from HeLa cells (HeLa NE , 20 ug). The molecular weight marker (in kDa) is shown on the left; the location of the protein of interest is indicated on the right.
Western blot analysis of LSD1 in A549 cell lysate with LSD1 antibody at 1 ug/mL.
Primary Antibodies
| Catalog No. | Host | Iso. | Clone | Pres. | React. | Applications | |
|---|---|---|---|---|---|---|---|
| AP08424PU-N | AOF2 (100-150) antibody |
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Rabbit | IgG | Aff - Purified | Can, Hu, Macaque, Mky, Ms, Rt, Ze | P, WB |
50 µg /
€425.00
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| OriGene Technologies GmbH | |||||||