EIF2AK3 antibody
Principal name
EIF2AK3 antibody
Alternative names for EIF2AK3 antibody
PEK, PERK, HsPEK, Eukaryotic translation initiation factor 2-alpha kinase 3, PRKR-like endoplasmic reticulum kinase, Pancreatic eIF2-alpha kinase
Ncbi ID
9606, NP_004827.4, 10090, NP_002437, NP_004827.3, NP_004827, NM_031599, NM_004836
Available reactivities
Available hosts
Available applications
Enzyme Immunoassay (E), Western blot / Immunoblot (WB), Immunocytochemistry/Immunofluorescence (ICC/IF), Paraffin Sections (P), Immunoprecipitation (IP)
Background of EIF2AK3 antibody
The PKR-like endoplasmic reticulum kinase (PERK, also know as Eukaryotic translation initiation factor 2-alpha kinase 3) is a type I transmembrane protein localized to the endoplasmic reticulum (ER). PERK consists of an N-terminal ER luminal domain, a membrane-spanning region, and a cytosolic C-terminal serine/threonine kinase domain (1). The luminal domain of PERK is bound to the ER chaperone GRP78 in unstressed cells (2). PERK activation occurs upon accumulation of misfolded proteins and the ER lumen, which triggers GRP78 dissociation from PERK thereby allowing PERK dimerization and autophosphorylation (3, 4). PERK phosphorylates two established targets: the eukaryotic translation initiation factor 2 alpha (eIF2alpha, (1)) and the Nrf2 transcription factor (5). Phosphorylation of eIF2alpha results in attenuation of translation initiation (6). The translational block also contributes to cell cycle arrest due to loss of the G1 regulatory protein, cyclin D1 (7). PERK-dependent phosphorylation of Nrf2 promotes transcription of phase II detoxifying enzymes which is critically important for elimination of intracellular reactive oxygen species (8). Thus, while inhibiting new protein synthesis and thereby decreasing the ER protein load PERK simultaneously induces expression of genes that help restore cellular redox homeostasis and promote survival.
EIF2AK3 polyclonal antibody (A01), Lot # 060525JCS1 Western Blot analysis of EIF2AK3 expression in PC-12 ( Cat # L012V1 ).
Immunohistochemistry analysis of PERK antibody (Cat.-No.: AP06757PU-N) on paraffin-embedded sections
Western blot (WB) analysis of PERK antibody (Cat.-No.: AP06757PU-N) in extracts from HeLa cells.
Formalin-fixed and paraffin-embedded human hepatocarcinoma tissue reacted with PERK antibody (N-term Q163), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining.
Immunohistochemistry staining of mouse mammary gland samples from lactating mice ( L10 ) with Eif2ak3 polyclonal antibody (Cat # PAB10175).Positive staining signal observed in wild type mouse sample with Eif2ak3 polyclonal antibody staining only ( B ) , but not in the knock out mouse sample ( C ) and pre-immune serum staining ( A ) . The Eif2ak3 polyclonal antibody ( Cat # PAB10175 ) was diluted in 1 : 1,000. Personal Communication. A, Diehl, Univ. of Pennsylvania, Philadelphia, PA.
Western blot analysis using Eif2ak3 polyclonal antibody ( Cat # PAB10175 ) to detect Eif2ak3 in cell lysates. 300 µg Eif2ak3 over-expressing 293T cell lysate ( lanes 1 & 2 ), or 800 µg wild type ( Lanes 3 & 4 ), and Eif2ak3 knock out ( lanes 5 & 6 ) MEF cell lysate were immunoprecipated with 15 µl anti-Eif2ak3, followed by immunobloting with 1:1000 dilution of Eif2ak3 polyclonal antibody in 5% milk/TBST buffer.Lane 1, 293T cells over-expressing Myc-Eif2ak3 wt, Lane 2 , 293T cells over-expressing Myc-Eif2ak3 K618A.Personal Communication. A, Diehl, Univ. of Pennsylvania, Philadelphia, PA.
Immunohistochemistry staining of Mouse mammary gland samples from lactating mice (L10) using AP09082SU-N PERK antibody. Positive staining signal observed in wild type Mouse sample with anti-PERK staining only (middle image), but not in the knock out mouse sample (Right image) and pre-immune serum staining (Left image). The anti-PERK was diluted 1/1,000 in 5% goat serum in PBS and allowed to incubate for 2h at RT in a humidified chamber. Personal Communication. A, Diehl, Univ. of Pennsylvania, Philadelphia, PA.
Western blot analysis: AP09082SU-N PERK antibody staining of cell lysates. 300µg PERK over-expressing 293T cell lysate (Lanes 1 and 2), or 800µg wild type (Lanes 3 and 4), and PERK knock out (Lanes 5 and 6) MEF cell lysate were immunoprecipated with 15µl anti-PERK, followed by immunobloting with 1/1000 dilution of anti-PERK in 5% milk/TBST buffer. Lane 1, 293T cells over-expressing Myc-PERK wt, Lane 2 , 293T cells over-expressing Myc-PERK K618A. Personal Communication. A, Diehl, Univ. of Pennsylvania, Philadelphia, PA.
Western blot analysis using EIF2AK3 antibody Cat.-No AM06529SU-N against HEK293 (1) and EIF2AK3 (AA: 929-1116)-hIgGFc transfected HEK293 (2) cell lysate.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue using PEK/PERK (Phospho-Thr981) antibody AP55771PU-N (left)or the same antibody preincubated with blocking peptide (right).
Immunohistochemical analysis of PERK staining in human brain formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Immunofluorescent analysis of PERK staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).
Western blot analysis of PERK expression in HeLa (A) whole cell lysates.
Immunoprecipitation of PERK from 0.5mg Hela whole cell extract lysate using 5ug of Anti-PERK Antibody and 50ul of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40ul SDS loading buffer and incubated for 10min at 70 C; 10ul of each sample was separated on a SDS PAGE gel transferred to a nitrocellulose membrane blocked with 5% BSA and probed with Anti-PERK Antibody.
Western blot analysis of extracts from MCF-7 cells, using PEK/PERK (Ab-981) antibody.The lane on the right is treated with the synthesized peptide.
Western blot analysis of extracts from JK cells (Lane 2), using PEK/PERK (Ab-981) Antibody. The lane on the left is treated with synthesized peptide.
Immunohistochemistry analysis of paraffin-embedded human colon carcinoma tissue using PEK/PERK (Ab-981) antibody.The picture on the right is treated with the synthesized peptide.
Western blot analysis of extracts from HeLa cells and HepG2 cells, using E2AK3 antibody.The lane on the right is treated with the synthesized peptide.
Immunofluorescence analysis of HepG2 cells, using E2AK3 antibody.The picture on the right is treated with the synthesized peptide.
Predicted cell location: Cytoplasm. Positive control: Human lung cancer tissue. Recommended dilution: 1/25-100 The image on the left is immunohistochemistry of paraffin-embedded human lung cancer tissue using EIF2AK3 antibody at dilution 1/30, on the right is treated with the fusion protein. (Original magnification:x200)
Predicted band size: 125kDa. Positive control: HT-29 cell lysate. Recommended dilution: 1/200-1000. (Gel: 8%SDS-PAGE Lysate:40 ug Primary antibody: 1/500 dilution Secondary antibody: Goat anti Rabbit IgG - H&L (HRP) at 1/8000 dilution Exposure time: 1 minute)
Predicted cell location: Cytoplasm. Positive control: Human liver cancer tissue. Recommended dilution: 1/50-200 The image on the left is immunohistochemistry of paraffin-embedded human liver cancer tissue using EIF2AK3 antibody at dilution 1/40, on the right is treated with the fusion protein. (Original magnification:x200)
293E cells were transfected with PERK-FLAG construct. Cell lysates were then immunoprecipitated with L5 antibody. Proteins were then treated with calf intestinal alkaline phosphatase (CIP). The proteins without CIP treatment were used as the control. Samples were separated by electrophoresis, transferred to nitrocellulose and probed with anti-PERK phospho (Ser713) antibody. Proteins were visualized using a donkey anti-rabbit secondary conjugated to HRP and a chemiluminenescent system.
Western blot analysis of PERK expression in Hela cells
Total cell lysate from 293T cells transfected with FLAG-PERK (lane 1) and FLAG-PERK S713A mutant (lane 2) were resolved by electrophoresis, transferred to nitrocellulose, and probed with purified anti-PERK Phospho (Ser713) antibody (clone Poly6494). Proteins were visualized using an HRP-conjugated anti-rabbit-IgG secondary and chemiluminescence detection.
The image on the left is immunohistochemistry of paraffin-embedded Human prostate cancer tissue using (EIF2AK3 Antibody) at dilution 1/50, on the right is treated with synthetic peptide. (Original magnification: ×200)
The image on the left is immunohistochemistry of paraffin-embedded Human brain tissue using (EIF2AK3 Antibody) at dilution 1/50, on the right is treated with synthetic peptide. (Original magnification: ×200)
Immunohistochemistry analysis of paraffin-embedded human prostate carcinoma tissue using PEK/PERK (Phospho-Thr981) antibody.The picture on the right is treated with the synthesized peptide.
Primary Antibodies
| Catalog No. | Host | Iso. | Clone | Pres. | React. | Applications | |
|---|---|---|---|---|---|---|---|
| AM06529SU-N | EIF2AK3 antibody |
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Mouse | IgG1 | 5G5 | Ascites | Hu | E, WB |
0.1 ml /
€380.00
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| OriGene Technologies GmbH | |||||||