The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Unit Definition: One unit is defined as the amount of Protein Kinase A regulatory subunit I alpha, required to incorporate 1nmol of phosphate into the specific substrate peptide kemptide (LRRASlG) in one minute at 30°C.
% SDS-PAGE. This protein was purified by proprietary chromatographic techniques.
This protein is cAMP free.
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 a 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.
For 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. Unit Definition: One unit is defined as the amount of Protein Kinase A regulatory subunit I alpha, required to incorporate 1nmol of phosphate into the specific substrate peptide kemptide (LRRASlG) in one minute at 30°C.
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Preparation and Storage
Stability and Storage
Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
Constituents: 50% Glycerol
This product is an active protein and may elicit a biological response in vivo, handle with caution.
cAMP dependent protein kinase regulatory subunit alpha 1
cAMP dependent protein kinase regulatory subunit RIalpha
cAMP dependent protein kinase type I alpha regulatory chain
cAMP dependent protein kinase type I alpha regulatory subunit
cAMP-dependent protein kinase type I-alpha regulatory subunit
Carney complex type 1
PRKAR1A/RARA FUSION GENE
Protein kinase A type 1a regulatory subunit
Protein kinase cAMP dependent regulatory type I alpha
PTC2 CHIMERIC ONCOGENE, INCLUDED
Tissue specific extinguisher 1
Tissue-specific extinguisher 1
Four types of regulatory chains are found: I-alpha, I-beta, II-alpha, and II-beta. Their expression varies among tissues and is in some cases constitutive and in others inducible.
Involvement in disease
Defects in PRKAR1A are the cause of Carney complex type 1 (CNC1) [MIM:160980]. CNC is a multiple neoplasia syndrome characterized by spotty skin pigmentation, cardiac and other myxomas, endocrine tumors, and psammomatous melanotic schwannomas. Defects in PRKAR1A are the cause of intracardiac myxoma (INTMYX) [MIM:255960]. Inheritance is autosomal recessive. Defects in PRKAR1A are the cause of primary pigmented nodular adrenocortical disease type 1 (PPNAD1) [MIM:610489]. Primary pigmented nodular adrenocortical disease is a rare bilateral adrenal defect causing ACTH-independent Cushing syndrome. Macroscopic appearance of the adrenals is characteristic with small pigmented micronodules observed in the cortex. PPNAD1 is most often diagnosed in patients with Carney complex, but it can also be observed in patients without other manifestations or familial history.
Belongs to the cAMP-dependent kinase regulatory chain family. Contains 2 cyclic nucleotide-binding domains.
The pseudophosphorylation site binds to the substrate-binding region of the catalytic chain, resulting in the inhibition of its activity.