Phospho-PKR (Thr446 + Thr451) Rabbit Polyclonal Antibody

• Catalog Number: orb6786
• Category: Antibodies
• Description: Phospho-PKR (Thr446 + Thr451) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Mouse, Rat
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthesised phosphopeptide derived from human PKR around the phosphorylation site of Thr446/451 KR(p-T)RSKG(p-T)LR
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=2ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 62 kDa
• Target: EIF2AK2
• UniProt ID: P19525
• RRID: AB_10927058
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: double-stranded RNA-dependent Protein Kinase; inte
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (black line): U251. Primary Antibody (green line): Rabbit Anti-Phospho-PKR (Thr446 + Thr451) antibody (orb6786), dilution: 2 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488, dilution: 0.5 ug/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°C, 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 20000 events was performed.

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) (orb6786) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 62 kDa, Observed band size: 60 kDa.

Sample: Liver (Mouse) Lysate at 40 ug, Primary: Anti-p-PKR (Thr446 + Thr451) (orb6786) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 62 kD, Observed band size: 62 kD.

Sample: Spleen (Mouse) Lysate at 40 ug, Primary: Anti-p-PKR (Thr446 + Thr451) (orb6786) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 62 kD, Observed band size: 62 kD.

Tissue/Cell: rat brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-Phospho-PKR (Thr446+Thr451) Polyclonal Antibody, Unconjugated (orb6786) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

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 (orb6786) 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.