Anti-Phospho-PKR/EIF2AK2-T446 Rabbit Polyclonal Antibody for WB, IF/ICC, ELISA - P19525

Core Product Specifications and Parameters

Biological Background: PKR/EIF2AK2 Function and Localization

• IFN-induced dsRNA-dependent serine/threonine-protein kinase that phosphorylates EIF2S1/eIF-2-alpha, leading to inhibition of global translation and playing a key role in innate immune response to viral infection. Related references: PMID:18835251 PMID:19189853 PMID:19507191

• Inhibits viral replication via the integrated stress response (ISR) by converting EIF2S1 into a global translational inhibitor and promoting ATF4 translation; exerts broad antiviral activity against DNA and RNA viruses including HCV, HBV, measles virus, and HSV-1. Related references: PMID:19189853 PMID:21123651 PMID:22948139 PMID:11836380 PMID:19840259

• Phosphorylates various substrates including p53/TP53, PPP2R5A, DHX9, ILF3, IRS1, and viral protein US11, thereby regulating signal transduction, apoptosis, cell proliferation, and differentiation. Related references: PMID:11836380 PMID:19229320 PMID:22214662

• Also functions as a tyrosine-protein kinase, phosphorylating CDK1 at Tyr-4 in response to DNA damage to promote its ubiquitination and proteasomal degradation. Related references: PMID:20395957

• Modulates multiple signaling pathways: activates NF-kappa-B via interaction with IKBKB and TRAF family proteins, activates p38 MAP kinase via MAP2K6, and regulates transcription factors (JUN, STAT1/3, IRF1, ATF3) involved in pro-inflammatory cytokine and IFN expression. Related references: PMID:22948139 PMID:23084476 PMID:23372823 PMID:10848580 PMID:15121867 PMID:15229216

• Acts as a dual regulator of the insulin signaling pathway: negatively via inhibitory phosphorylation of IRS1 at Ser-312, and positively by phosphorylating PPP2R5A, which activates FOXO1 and up-regulates IRS2. Related references: PMID:20685959

• Regulates assembly and activation of NLRP3, NLRP1, AIM2, and NLRC4 inflammasomes. Related references: PMID:22801494

• Predominantly cytoplasmic with nuclear and perinuclear localization; highly expressed in thymus, spleen, and bone marrow; elevated levels in breast and colon carcinomas correlate with tumor progression; colocalizes with GSK3B and TAU in Alzheimer’s disease brain.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phosphopeptide around Thr446; phosphatase inhibitors in lysis buffers are critical to preserve phosphorylation during sample preparation.
• For western blotting, validate phospho-specificity by comparing signals from phosphatase-treated versus untreated lysates or using a non-phospho peptide blocking control.
• In immunofluorescence, consider antigen retrieval conditions and include a nuclear counterstain to visualize the cytoplasmic/perinuclear distribution.
• When applying ELISA, pair with a total PKR antibody to normalize for protein abundance and assess phosphorylation ratios.
• Cross-reactivity is reported for human and mouse; verify performance in other species before use.

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