Application Note
Protein Kinase R (PKR) is activated by double-stranded RNA (dsRNA), the synthesis of which is caused virally. PKR can also be activated by the protein PACT or by heparin. It plays a key role in the innate immune response to viral infection and is also involved in the regulation of signal transduction, apoptosis, cell proliferation and differentiation. Besides, Cyclin Dependent Kinase 1 (CDK1) has been identified as an interactor of PKR, thus a binding ELISA assay was conducted to detect the interaction of recombinant human PKR and recombinant human CDK1. Briefly, PKR were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to CDK1-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-PKR pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of of PKR and CDK1 was in a dose dependent manner.
Observed MW
32 kDa.
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Ser224~Ile502 (NP_001129123.1)
Expression System
E. coli
Purity
> 80%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Synonyms
eukaryotic translation initiation factor 2 alpha kinase 2 , EIF2AK1 , PKR , PPP1R83 , PRKR
Background
The protein encoded by this gene is a serine/threonine protein kinase that is activated by autophosphorylation after binding to dsRNA. The activated form of the encoded protein can phosphorylate translation initiation factor EIF2S1, which in turn inhibits protein synthesis. This protein is also activated by manganese ions and heparin. Three transcript variants encoding two different isoforms have been found for this gene. [provided by RefSeq, Oct 2011]
Database
Research Area