Human Protein Kinase R (PKR) Protein

Human Protein Kinase R (PKR) Protein (Active) is a recombinant active Human protein expressed in E. coli.

Target	Protein Kinase R (PKR)
Origin	Human
Tested Applications	WB, SDS-PAGE
Host	E. coli
Conjugation	Unconjugated
Form	Lyophilized
Purity	> 80%
Reconstitution	Reconstitute in ddH₂O to a concentration of 0.1-1.0 mg/ml. Do not vortex.
Storage	Store at 2-8 °C for up to one month. Store at -80 °C for up to one year. Avoid repeated freeze/thaw cycles.
UniProt Primary AC	P19525 (UniProt, ExPASy)
UniProt Secondary AC	A8K3P0, D6W584, E9PC80, Q52M43, Q7Z6F6, Q9UIR4
UniProt Entry Name	E2AK2_HUMAN
KEGG	hsa:5610
String	9606.ENSP00000233057
Molecular Weight	Calculated MW: 35.8 kDa Observed MW: 32 kDa Possible reasons why the actual band size differs from the predicted band size: 1. Splice variants. Alternative splicing may create different sized proteins from the same gene. 2. Relative charge. The composition of amino acids may affect the charge of the protein. 3. Post-translational modification. Phosphorylation, glycoslyation, methylation etc. may affect the band size. 4. Post-translational cleavage. Many proteins are synthesised as pro-proteins, and then cleaved to give the active form. 5. Polymerisation of the target protein. Dimerisation, multimerisation etc. will increase the band size observed.
Sequence Fragment	Ser224-Ile502
Sequence	SSSLLMN GLRNNQRKAK RSLAPRFDLP DMKETKYTVD KRFGMDFKEI ELIGSGGFGQ VFKAKHRIDG KTYVIKRVKY NNEKAEREVK ALAKLDHVNI VHYNGCWDGF DYDPETSDDS LESSDYDPEN SKNSSRSKTK CLFIQMEFCD KGTLEQWIEK RRGEKLDKVL ALELFEQITK GVDYIHSKKL IHRDLKPSNI FLVDTKQVKI GDFGLVTSLK NDGKRTRSKG TLRYMSPEQI SSQDYGKEVD LYALGLILAE LLHVCDTAFE TSKFFTDLRD GI
Tag	N-terminal His tag
Buffer	Prior to lyophilization: 20 mM Tris, 150 mM NaCl, pH 8.0, containing 0.01% Sarcosyl, 5% Trehalose.
Activity	Active
Biological Activity	Protein Kinase R (PKR) is activated by double-stranded RNA (dsRNA), the synthesis of which is caused by viruses. PKR can also be activated by the protein PACT or by heparain. 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. 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 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to CDK1-coated microplate wells and incubated for 2 h at 37°C. Wells were washed with PBST and incubated for 1 h with anti-PKR polyclonal antibody, then aspirated and washed 3 times. After incubation with HRP-conjugated secondary antibody, wells were aspirated and washed 3 times. TMB substrate solution was added and wells were incubated for 15-25 minutes at 37 °C. Finally, 50 μl stop solution was added to the wells and the absorbance was read at 450 nm immediately. The binding activity of PKR and CDK1 is shown in Figure 4.
Endotoxin Level	< 1.0 EU/µg (LAL method)
Concentration	Prior to lyophilization: 200 µg/ml
Availability	Shipped within 5-7 working days.
Note	This product is for research use only. Not for human consumption, cosmetic, therapeutic or diagnostic use.

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Human Protein Kinase R (PKR) Protein

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