Human PRKACA Recombinant Protein Product Attributes
Product Type: Recombinant Protein
Recombinant PRKACA based upon sequence from Human
Host: QP13140 protein expressed in E. Coli, Insect.
Available Tags: Untagged, GST
Protein Construction: A cDNA sequence encoding the sequence of PRKACA was constructred and used to recombinantly synthesize the protein.
Recommended Applications: See Application Notes
Application Notes
Definition of Unit (for specific activity): One unit is defined as the amount of cAMP-Dependent Protein Kinase, recombinant C? catalytic subunit, required to incorporate 1 pmol of phosphate into the specific substrate peptide kemptide (LRRASLG) in one minute at 30°C.
Other Applications: Roskoski-Assay Protein kinase activity can be measured using a modified radioactive assay according to Roskoski et al. The assay is performed in a mixture of50mM 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.
Bioactivity Data:
Specific Activity: The specific activity of the recombinant PKA catalytic subunit alpha, is >10,000,000 U/mg.
Amino Acid Sequence: MGNAAAAKKG SEQESVKEFL AKAKEDFLKK WESPAQNTAH LDQFERIKTL GTGSFGRVML VKHKETGNHY AMKILDKQKV VKLKQIEHTL NEKRILQAVN FPFLVKLEFS FKDNSNLYMV MEYVPGGEMF SHLRRIGRFS EPHARFYAAQ IVLTFEYLHS LDLIYRDLKP ENLLIDQQGY IQVTDFGFAK RVKGRTWTLC GTPEYLAPEI ILSKGYNKAV DWWALGVLIY EMAAGYPPFF ADQPIQIYEK IVSGKVRFPS HFSSDLKDLL RNLLQVDLTK RFGNLKNGVN DIKNHKWFAT TDWIAIYQRK VEAPFIPKFK GPGDTSNFDD YEEEEIRVSI NEKCGKEFSE F
Purity: Greater than 95% as determined by SDS-PAGE.
Reconstitution Instructions: |||
Endotoxin Levels: Not Determined. Testing recommended prior to use in cell culture and can be performed upon request.
Buffer: PKA catalytic subunit a is supplied in a buffer containing 20mM MOPS pH 7, 150mM NaCl, 1mM DTT, 1mM EDTA and 50% Glycerin.
Storage Conditions: Store at 4°C if entire vial will be used within 2-4 weeks. Store, frozen at -20°C for longer periods of time. Avoid multiple freeze-thaw cycles.
Physical Appearance: Sterile Colorless Liquid
Limitations and Performance Guarantee
enQuire Bio's products are guaranteed and approved for RESEARCH USE ONLY or manufacturing purposes. This product may not be used as a pharmaceutical agent, or other product intended for human consumption without meeting additional regulatory requirements. This product is guaranteed to work for a period of two years when stored at -70C or colder, and one year when aliquoted and stored at -20C.
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