Cell Signaling Technology

Product Pathways - Translational Control

PKR Antibody #3072

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC H Endogenous 74 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

PKR Antibody detects endogenous levels of PKR. This antibody does not cross-react with other related proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr451 of human PKR. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or IFN-alpha/beta-treated (1000 U/ml for 18 hours), and subsequently untreated or calyculin A-treated ( 0.1 µM for 15 minutes), using PKR Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or IFNalpha-treated (18 hrs., right), using PKR Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

Background

Protein kinase R (PKR) is transcriptionally induced by interferon and activated by double-stranded RNA (dsRNA). PKR inhibits translation initiation through phosphorylation of the alpha subunit of the initiation factor eIF2 (eIF2alpha) and also controls the activation of several transcription factors such as NF-kappaB, p53 and the Stats. In addition, PKR mediates apoptosis induced by many different stimuli, such as LPS, TNF-alpha, viral infection and serum starvation (1,2). Activation of PKR by dsRNA results in PKR dimerization and autophosphorylation at threonine 446 and threonine 451 in the activation loop. Substitution of threonine 451 to alanine completely inactivates PKR, while a mutant with a threonine 446 to alanine substitution was partially active (3). Recently, activation of PKR has been implicated in the pathologies of neurodegenerative diseases, including Alzheimer disease (4,5).

  1. Williams, B.R. (1999) Oncogene 18, 6112-6120.
  2. Gil, J. and Esteban, M. (2000) Apoptosis 5, 107-114.
  3. Romano, P. R. et al. (1998) Mol. Cell. Biol. 18, 2282-2297.
  4. Peel, A.L. and Bredesen, D.E. (2003) Neurobiol. Dis. 14, 52-62.
  5. Peel, A.L. (2004) J. Neuropath. Exp. Neurol. 63, 97-105.

Application References

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Companion Products

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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