Datasheet Version: 3.0.0 Revision date: 23 Apr 2021 Human Protein Kinase R (PKR) Protein (Active) Catalogue No.:abx651433 SDS-PAGE analysis of recombinant Human PKR. Western blot analysis of recombinant Human PKR, using PKR antibody (abx128520). Gene sequencing extract of recombinant Human PKR. Binding activity of PKR with CDK1. For Reference Only 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 v1.0.0 Abbexa LTD, Cambridge, UK · Phone: +44 (0) 1223 755950 · Fax: +44 (0) 1223 755951 1 of 3 Abbexa LLC, Houston, TX USA · Phone: +1 832 327 7413 Website: www.abbexa.com · Email: [email protected] Datasheet Version: 3.0.0 Revision date: 23 Apr 2021 Host: E. coli Conjugation: Unconjugated Form: Lyophilized Purity: > 80% Reconstitution: Reconstitute to the original concentration in ddH2O. If further dilutions are required, dilute in 20 mM Tris, 150 mM NaCl, pH 8.0, 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) 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 K ForTYVIKRVKY NNEKAEREVKReference ALAKLDHVNI VHYNGCWDGF DYDPETSDDS Only LESSDYDPEN SKNSSRSKTK CLFIQMEFCD KGTLEQWIEK RRGEKLDKVL ALELFEQITK GVDYIHSKKL IHRDLKPSNI FLVDTKQV KI GDFGLVTSLK NDGKRTRSKG TLRYMSPEQI SSQDYGKEVD LYALGLILAE LLHVCDTAFE TSKFFT DLRD GI Tag: N-terminal His-tag Buffer: Prior to lyophilization: 20 mM Tris, 150 mM NaCl, pH 8.0, containing 1 mM EDTA, 1 mM DTT, 0.01% Sarcosyl, 5% Trehalose and Proclin-300. v1.0.0 Abbexa LTD, Cambridge, UK · Phone: +44 (0) 1223 755950 · Fax: +44 (0) 1223 755951 2 of 3 Abbexa LLC, Houston, TX USA · Phone: +1 832 327 7413 Website: www.abbexa.com · Email: [email protected] Datasheet Version: 3.0.0 Revision date: 23 Apr 2021 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 Note: This product is for research use only. Not for human consumption, cosmetic, therapeutic or diagnostic use. For Reference Only v1.0.0 Abbexa LTD, Cambridge, UK · Phone: +44 (0) 1223 755950 · Fax: +44 (0) 1223 755951 3 of 3 Abbexa LLC, Houston, TX USA · Phone: +1 832 327 7413 Website: www.abbexa.com · Email: [email protected].