Anti-Phospho-PKR (Thr446+Thr451) Polyclonal Antibody is a Rabbit antibody targeting Phospho-PKR (Thr446+Thr451). Anti-Phospho-PKR (Thr446+Thr451) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human,Mouse,Rat

1. Sample:Spleen (Mouse) Lysate at 40 μg
Primary: Anti-p-PKR (Thr446+Thr451) (TMAB-11161) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 62 kD
Observed band size: 62 kD
2. Sample: Liver (Mouse) Lysate at 40 μg
Primary: Anti-p-PKR (Thr446+Thr451) (TMAB-11161) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 62 kD
Observed band size: 62 kD
3. Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-Phospho-PKR(Thr446+Thr451) Polyclonal Antibody, Unconjugated (TMAB-11161) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
4. U251 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Phospho-PKR (Thr446+Thr451)) polyclonal Antibody, Unconjugated (TMAB-11161) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
5. Sample:
Lane 1: Human HeLa cell lysates
Lane 2: Human K562 cell lysates
Lane 3: Human 293T cell lysates
Lane 4: Human A431 cell lysates
Primary: Anti-Phospho-PKR (Thr446+Thr451) (TMAB-11161) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 62 kDa
Observed band size: 60 kDa
6. Blank control (black line): U251.
Primary Antibody (green line): Rabbit Anti-Phospho-PKR (Thr446+Thr451) antibody (TMAB-11161)
Dilution: 2 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min 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.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100-500; IF: 1:100-500; FCM: 2μg/Test

Polyclonal

KLH conjugated synthesised phosphopeptide: human PKR around the phosphorylation site of Thr446/451

Human

EIF2AK2

Interferon-induced, double-stranded RNA-activated protein kinase

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.