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Merck

Measurement of RNA-induced PKR Activation in vitro.

Bio-protocol (2017-03-20)
Kobe C Yuen
ABSTRACT

Protein kinase R (PKR) is one of the key RNA-activated sensors for innate immunity. PKR is activated by pathogenic or aberrant RNAs such as short double-stranded RNAs or those with imperfect secondary structures, as well as a reduction in the amount and number of RNA modifications. Activation of PKR may be an underlying mechanism for the pathogenesis of human diseases. In this protocol, I describe a method for studying levels of RNA-induced PKR activation in vitro.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Phenylmethanesulfonyl fluoride, ≥98.5% (GC)
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Ribonuclease A from bovine pancreas, (Solution of 50% glycerol, 10mM Tris-HCL pH 8.0)
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Sodium orthovanadate, ≥90% (titration)
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Sodium chloride, BioXtra, ≥99.5% (AT)
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HEPES, ≥99.5% (titration)
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Imidazole, Molecular Biology, ≥99% (titration)
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DL-Dithiothreitol, ≥98% (HPLC), ≥99.0% (titration)
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TWEEN® 20, Molecular Biology, viscous liquid
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Potassium chloride, BioXtra, ≥99.0%
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Bovine Serum Albumin, lyophilized powder, BioReagent, suitable for cell culture
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Sodium hydroxide, BioXtra, ≥98% (acidimetric), pellets (anhydrous)
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Ethylenediaminetetraacetic acid, anhydrous, crystalline, BioReagent, suitable for cell culture
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Sodium phosphate monobasic, BioXtra, ≥99.0%
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IPTG, ≥99% (TLC), ≤0.1% Dioxane
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Magnesium chloride, ≥98%
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