Phospho-PKR (Thr446 + Thr451) Rabbit Polyclonal Antibody

SKU: orb6786

Description

Phospho-PKR (Thr446 + Thr451) Rabbit Polyclonal Antibody

Images & Validation

• Tested Applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100-500, IF=1:100-500, Flow-Cyt=2ug/Test
• Reactivity: Human, Mouse, Rat
• Predicted Reactivity: Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• 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
• Target: EIF2AK2
• Molecular Weight: 62 kDa
• Purification: Affinity purified by Protein A

Storage & Handling

• 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.
• Form/Appearance: Liquid
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Concentration: 1mg/ml
• Disclaimer: For research use only

Alternative Names

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 activated protein kinase; p68 kinase; eIF-2A protein kinase 2; P1/eIF-2A protein kinase; protein kinase RNA-activated; interferon-inducible RNA-dependent protein kinase; EIF2AK2; EIF2AK1; MGC126524; PKR; PRKR; E2AK2_HUMAN.

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.

Quick Database Links

Gene Symbol

EIF2AK2

UniProt

P19525