Phospho-PKR (T446) Recombinant Rabbit Monoclonal Antibody [SY230] - BSA and Azide free
Catalog# HA750113
Phospho-PKR (T446) Recombinant Rabbit Monoclonal Antibody [SY230] - BSA and Azide free
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WB
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IHC-P
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IP
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IF-Cell
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Human
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ET1607-20
含抗保成分
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unconjugated
Overview
Product Name
Phospho-PKR (T446) Recombinant Rabbit Monoclonal Antibody [SY230] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Thr446 of Human PKR aa 421-470 / 551.
Species Reactivity
Human
Validated Applications
WB, IHC-P, IP, IF-Cell
Molecular Weight
Predicted band size: 62 kDa
Positive Control
Jurkat treated with 100nM Calyculin A for 30 minutes cell lysate, Hela treated with Calyculin A and TNF-alpha whole cell lysate, human tonsil tissue, human spleen tissue, human breast tissue, human breast carcinoma tissue, human kidney tissue, human small intestine tissue, SiHa.
Conjugation
unconjugated
Clone Number
SY230
Product Features
Form
Liquid
Concentration
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:5,000
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IHC-P
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1:50-1:200
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IP
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Use at an assay dependent concentration.
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IF-Cell
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1:50
Target
Function
An interferon-inducible, RNA-dependent protein serine/threonine kinase, PKR has various designations. Mouse PKR is known as DAI, dsJ, PI kinase, p65, p67 or TIK, whereas human PKR is known as p68 or p69. PKR phosphorylates its substrate, a subunit of protein synthesis initiation factor eIF-2 on Ser 51 to inhibit translation. PKR contains two dsRNA binding motifs required for its activation by dsRNA. Three kinds of regulation of PKR enzymatic activity occur, and these include transcriptional regulation in response to interferon, an autoregulatory mechanism controlling PKR expression at the level of translation, and posttranslational regulation by RNA mediated autophosphorylation. Human PKR contains at least 15 autophosphorylation sites, but only Thr-446 and Thr-451 in the activation loop are critical for its kinase activity. Thr-446 is the in vivo autophosphorylation site of PKR. Mutation of threonine to alanine at position 446 substantially reduces PKR function, and mutant kinase containing Ala-451 is completely inactive.
Background References
1. Pfaller CK et al. Measles virus C protein impairs production of defective copyback double-stranded viral RNA and activation of protein kinase R. J Virol 88:456-68 (2014).
2. Nejepinska J et al. Reporters transiently transfected into mammalian cells are highly sensitive to translational repression induced by dsRNA expression. PLoS One 9:e87517 (2014).
Sequence Similarity
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. GCN2 subfamily.
Tissue Specificity
Highly expressed in thymus, spleen and bone marrow compared to non-hematopoietic tissues such as small intestine, liver, or kidney tissues. Colocalizes with GSK3B and TAU in the Alzheimer disease (AD) brain. Elevated levels seen in breast and colon carcinomas, and which correlates with tumor progression and invasiveness or risk of progression.
Post-translational Modification
Autophosphorylated on several Ser, Thr and Tyr residues. Autophosphorylation of Thr-451 is dependent on Thr-446 and is stimulated by dsRNA binding and dimerization. Autophosphorylation apparently leads to the activation of the kinase. Tyrosine autophosphorylation is essential for efficient dsRNA-binding, dimerization, and kinase activation.
Subcellular Location
Cytoplasm, Nucleus.
UNIPROT
Synonyms
Double stranded RNA activated protein kinase
antibody
E2AK2_HUMAN antibody
eIF-2A protein kinase 2 antibody
EIF2AK1 antibody
EIF2AK2 antibody
Eukaryotic translation initiation factor 2 alpha kinase 2 antibody
Eukaryotic translation initiation factor 2-alpha kinase 2 antibody
HGNC:9437 antibody
Interferon induced double stranded RNA activated protein kinase antibody
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of Phospho-PKR (T446) on different lysates with Rabbit anti-Phospho-PKR (T446) antibody (HA750113) at 1/5,000 dilution.
Lane 1: Jurkat cell lysate
Lane 2: Jurkat treated with 100nM Calyculin A for 30 minutes cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 62 kDa
Observed band size: 68 kDa
Exposure time: 30 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750113) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Phospho-PKR (T446) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750113, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Phospho-PKR (T446) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750113, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-Phospho-PKR (T446) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750113, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-PKR (T446) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750113, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Phospho-PKR (T446) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750113, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-Phospho-PKR (T446) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750113, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunocytochemistry analysis of SiHa cells labeling Phospho-PKR (T446) with Rabbit anti-Phospho-PKR (T446) antibody (HA750113) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-PKR (T446) antibody (HA750113) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Products with the same target and pathway
PKR Recombinant Rabbit Monoclonal Antibody [SC06-37]
Application: WB
Reactivity: Human
Conjugate: unconjugated
Phospho-PKR (T451) Recombinant Rabbit Monoclonal Antibody [JE46-70]
Application: WB
Reactivity: Human
Conjugate: unconjugated
