PKA [GST-tagged]

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Species Reactivity: Human  

Source: E. coli expression  

Formula: 50 mM Tris/HCl pH 7.5, 0.1 mM EGTA, 150 mM NaCl, 270 mM sucrose, 0.03% Brij, 0.1% b-Mercaptoethanol, 1 mM Benzamidine, 0.2 mM PMSF  

Concentration: 1 mg/ml  

Molecular Weight: ~67.9 kDa  

Protein Sequence: Accession number: NP_002721.1. For full protein sequence information download the Certificate of Analysis pdf.  

Quality Control Protein Identification: Confirmed by mass spectrometry.  

Quality Control Activity: Activity Assay:The specific activity of GST-PKA was determined using the method described by Hastie et al. (2006) with the enzyme being assayed at several concentrations.   GST-PKA was incubated for 10 minutes at 30 °C in kinase reaction buffer in the presence of KEMPtide substrate (30 µM)   and [γ-32P]ATP (100 µM). Duplicate reactions were stopped by spotting the assay mixture onto Whatman P81 paper - capturing the phosphorylated substrate.   The radioactivity incorporated was measured on a scintillation counter and the enzyme’s mean specific activity was calculated.  GST-PKA specific activity:1394.0 Units/mg (1394.0 Units/ml)1 Unit = 1 nmole of phosphate incorporated into the substrate in 1 minuteSubstrate: KEMPtide (LRRASLG)