Rabbit Anti-EIF2AK2 (PKR) antibody

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double-stranded RNA-dependent Protein Kinase; interferon-induced, double-stranded RNA-activated protein kinase isoform a; protein kinase, interferon-inducible double stranded RNA dependent; interferon-inducible elF2alpha kinase; double stranded RNA activa

Cat:

SL1493R

Species Reactivity：

Human,Mouse,Rat,

Immunogen：

KLH conjugated synthetic peptide derived from human PKR:251-360/551

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=1ug/testIF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: Hela(Human) Cell Lysate at 30 ug293T(Human) Cell Lysate at 30 ugPrimary: Anti-PKR (SL1493R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 61 kDObserved band size: 61 kDSample: Liver (Mouse) Lysate at 40 ugPrimary: Anti- PKR (SL1493R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 61 kDObserved band size: 61 kDSample:Spleen (Mouse) Lysate at 40 ugPrimary: Anti- PKR (SL1493R)at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 61 kDObserved band size: 61 kD Paraformaldehyde-fixed, paraffin embedded ( rat spleen tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PKR) Polyclonal Antibody, Unconjugated (SL1493R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining. Paraformaldehyde-fixed, paraffin embedded (rat kidney tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (EIF2AK2) Polyclonal Antibody, Unconjugated (SL1493R) at 1:200 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining. Tissue/cell: rat kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-EIF2AK2/PKR Polyclonal Antibody, Unconjugated(SL1493R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingTissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-EIF2AK2/PKR Polyclonal Antibody, Unconjugated(SL1493R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingA431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (PKR) polyclonal Antibody, Unconjugated (SL1493R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.Blank control: Raji. Primary Antibody (green line): Rabbit Anti-PKR antibody (SL1493R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PEDilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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Unit:

Price: $220.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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PKR is an interferon-inducible serine/threonine specific protein kinase. It is widely expressed in eukaryotic organisms and activated by double stranded RNA. Activation of PKR by dsRNAs leads to autophosphorylation at multiple sites. Phosphorylation of Thr446 and Thr451 in the PKR activation loop is required in vivo and in vitro for high level kinase activity. PKR phosphorylates its natural substrate, the alpha subunit of eukaryotic protein synthesis initiation factor 2 (EIF2 alpha), leading to the inhibition of protein synthesis. PKR is also involved in TLR signaling and mediates apoptosis in fibroblasts in response to viral infection and inflammatory cytokines, and also activates IKK and NFKB, thereby suppressing apoptosis. Recently, it has been reported that PKR also phosphorylates human p53 on serine 392. PKR might play a role in ER stress-induced apoptosis and in Alzheimer's disease. Alzheimer cases show prominent PKR activation in association with neuritic plaques and pyramidal neurons in the hippocampus and neocortex.

Function:
Following activation by double-stranded RNA in the presence of ATP, the kinase becomes autophosphorylated and can catalyze the phosphorylation of the translation initiation factor EIF2S1, which leads to an inhibition of the initiation of protein synthesis. Double-stranded RNA is generated during the course of a viral infection.

Subunit:
Homodimer. Interacts with STRBP. Interacts with DNAJC3. Inhibited by direct interaction with viral proteins such as HCV E2, HCV NS5A and influenza A NS1. Activated by the interaction with HISLV1 Tat. Forms a complex with FANCA, FANCC, FANCG and HSP70.

Post-translational modifications:
Autophosphorylated on several Ser and Thr 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.

Similarity:
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. GCN2 subfamily.
Contains 2 DRBM (double-stranded RNA-binding) domains.
Contains 1 protein kinase domain.

SWISS:
P19525

Gene ID:
5610

Database links:

Entrez Gene: 5610 Human

Omim: 176871 Human

SwissProt: P19525 Human

Unigene: 131431 Human

蛋白激酶R/双链RNA依赖蛋白激酶（PKR）是一种干扰素诱导的、双链RNA激活的丝氨酸/苏氨酸激酶, PKR在信号转导、细胞生长、分化和凋亡的控制中起重要作用.也有人认为：PKR是一个重要的凋亡效应物,是许多不同刺激物诱导凋亡的一个转导物。 Picture

Sample:
Liver (Mouse) Lysate at 40 ug
Primary: Anti- PKR (SL1493R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 61 kD
Observed band size: 61 kD

Sample:
Spleen (Mouse) Lysate at 40 ug
Primary: Anti- PKR (SL1493R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 61 kD
Observed band size: 61 kD

Paraformaldehyde-fixed, paraffin embedded ( rat spleen tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PKR) Polyclonal Antibody, Unconjugated (SL1493R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat kidney tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (EIF2AK2) Polyclonal Antibody, Unconjugated (SL1493R) at 1:200 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.

Tissue/cell: rat kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-EIF2AK2/PKR Polyclonal Antibody, Unconjugated(SL1493R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining

Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-EIF2AK2/PKR Polyclonal Antibody, Unconjugated(SL1493R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining

A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (PKR) polyclonal Antibody, Unconjugated (SL1493R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Blank control: Raji.
Primary Antibody (green line): Rabbit Anti-PKR antibody (SL1493R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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