Phospho-PKR (T446) Recombinant Rabbit Monoclonal Antibody [SY230]
cat.: ET1607-20
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IHC-P, IP, IF-Cell
Clonality: Monoclonal
Clone number: SY230
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 62 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Thr446 of Human PKR aa 421-470 / 551.
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.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  IP
  IF-Cell
1:500-1:5,000
1:50-1:200
Use at an assay dependent concentration.
1:50
Uniprot #: SwissProt: P19525 Human
Alternative names: Double stranded RNA activated protein kinase; E2AK2_HUMAN eIF-2A protein kinase 2 EIF2AK1 EIF2AK2 Eukaryotic translation initiation factor 2 alpha kinase 2 Eukaryotic translation initiation factor 2-alpha kinase 2 HGNC:9437 Interferon induced double stranded RNA activated protein kinase Interferon inducible elF2 alpha kinase Interferon inducible RNA dependent protein kinase Interferon-induced, double-stranded RNA-activated protein kinase Interferon-inducible RNA-dependent protein kinase MGC126524 P1/eIF-2A protein kinase P1/eIF2A protein kinase p68 kinase PKR PPP1R83 PRKR Protein kinase interferon inducible double stranded RNA dependent Protein kinase RNA activated Protein kinase RNA-activated Protein phosphatase 1 regulatory subunit 83 Serine/threonine protein kinase TIK Tyrosine protein kinase EIF2AK2
Images
ET1607-20_1.jpg Fig1: Western blot analysis of Phospho-PKR (T446) on different lysates with Rabbit anti-Phospho-PKR (T446) antibody (ET1607-20) 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 (ET1607-20) 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.
ET1607-20_2.jpg Fig2: Western blot analysis of Phospho-PKR (T446) on different lysates using anti-Phospho-PKR (T446) antibody at 1/500 dilution.
Lane 1: Hela treated with Calyculin A and TNF-alpha whole cell lysates
Lane 2: Untreated Hela whole cell lysates
ET1607-20_3.jpg Fig3: 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) for 20 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 (ET1607-20, 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.
ET1607-20_4.jpg Fig4: 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) for 20 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 (ET1607-20, 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.
ET1607-20_5.jpg Fig5: 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) for 20 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 (ET1607-20, 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.
ET1607-20_6.jpg Fig6: 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) for 20 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 (ET1607-20, 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.
ET1607-20_7.jpg Fig7: 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) for 20 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 (ET1607-20, 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.
ET1607-20_8.jpg Fig8: 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) for 20 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 (ET1607-20, 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.
ET1607-20_9.jpg Fig9: Immunocytochemistry analysis of SiHa cells labeling Phospho-PKR (T446) with Rabbit anti-Phospho-PKR (T446) antibody (ET1607-20) 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 (ET1607-20) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.