ANTI-PHOSPHO-PKR (PTHR451) HUMAN DEVELOPED IN RABBIT AFFINITY ISOLATED ANTIBODY

General description

Protein kinase regulated by RNA (PKR) is an interferon-inducible ser/thr protein kinase, activated by stress signals and double-stranded RNA (dsRNA). In response to viral infection, PKR is activated by dsRNA-mediated dimerization and autophosphorylation. α-subunit of protein synthesis initiation factor 2 (eIF-2α) is well-studied substrate of PKR. Activation by PKR leads to phosphorylation on Ser51 that result in inhibition of translation. Human PKR contains at least 15 autophosphorylation sites. But phosphorylation on threonine 446 and 451 in the PKR activation loop is required ^{in vivo} and in vitro for high-level kinase activity. PKR is implicated in signalling pathways that involve NF-κB, p38, and IRF-3 and is the key effector in IFN-induced anti-viral responses, apoptosis and protein synthesis.
Anti-Phospho-PKR [pThr⁴⁵¹] specifically recognizes PKR phosphorylated on threonine 451 (65-68 kDa). The antibody detects human and mouse PKR [pThr⁴⁵¹].

Immunogen

synthetic phosphopeptide derived from the region of PKR that is phosphorylated on threonine 451.

Application

Anti-phospho-PKR (pThr⁴⁵¹) antibody may be used for detection by immunoblotting at a working dilution of 1:1,000 using human HeLa cells stimulated with IFN·γ and calyculin. For detection by immunoblotting in human HeLa cells, a working concentration of 0.1-1.0 μg/mL is recommended. Detection of phospho-PKR (pThr⁴⁵¹) was done in peritoneal macrophages obtained from thioglycolate-treated mice and human peripheral blood mononuclear cells at a working dilution of 1:500

Physical form

Supplied as a solution in Dulbecco′s phosphate buffered saline (without Mg²⁺ and Ca²⁺), pH 7.3 (+/- 0.1), 50% glycerol, with 1.0 mg/mL BSA (IgG, protease free) as a carrier and 0.05% sodium azide. The amount of the reagent is sufficient for 10 blots.

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