Recombinant Human cAMP Protein Kinase Catalytic Subunit (A63047)

Lot Specific

Supplied in 20 mM MOPS Buffer, pH 7, with 150 mM Sodium Chloride, 1 mM DTT, 1 mM EDT, and 50% Glycerin.

Shipped at 4°C. Short term (2-4 weeks) store at 4°C. Long term store at -20°C. Avoid freeze/thaw cycles.

Roskoski-AssayProtein kinase activity can be measured using a modified radioactive assay according to Roskoski et al.The assay will be performed in a mixture containing 50mM MOPS (pH7.0), 10mM MgCI2, 0.25 mg/ml bovine serum albumin, 100 IJM Kemptide (peptide substrate), 100 IJM unlabeled ATP mixed with [y_32p] ATP (500-1000 cpm/pmol) and Ca subunit in a final volume of 50 IJI. Reaction is started by addition of the Ca subunit and can be stopped after 5 minutes incubation at 30°C by spotting the reaction mix onto Whatman P-81 filters and soaking the filters four times in 75mM phosphoric acid (10 ml per sample) for at least 5 minutes. After four washing steps rinse filters with ethanol, dry and count. Roskoski, R., Jr. (1983) Methods Enzymol. 99, 3-6For the detection of phosphorylation in substrate proteins the phosphotransferase reaction can alternatively be stopped by taking aliquots of the mixture and adding SDS sample buffer. The phosphorylation status of the substrate proteins can subsequently be analysed using SDS PAGE and autoradiography. Zimmermann, B. (1999) Journal of Biological Chemistry.274, 9, 5370-78.

cAMP dependent protein kinase alpha catalytic subunit, cAMP dependent protein kinase beta catalytic subunit, cAMP dependent protein kinase catalytic beta subunit isoform 4ab, cAMP dependent protein kinase catalytic subunit alpha, cAMP dependent protein kinase catalytic subunit alpha, isoform 1, cAMP dependent protein kinase catalytic subunit beta, cAMP-dependent protein kinase catalytic subunit alpha, KAPCA_HUMAN, PKA C alpha, PKA C beta, PKA C-alpha, PKACA, PKACB, PPNAD4, PRKACA, PRKACAA, PRKACB, Protein kinase A catalytic subunit, Protein kinase A catalytic subunit alpha, Protein kinase A catalytic subunit beta, Protein kinase, cAMP dependent, catalytic, alpha, Protein kinase, cAMP dependent, catalytic, beta