The second messenger cyclic AMP (cAMP) activates cAMP-dependent protein kinase (PKA or cAPK) in mammalian cells and controls many cellular mechanisms such as gene transcription, ion transport, and protein phosphorylation (1).
Inactive PKA is a heterotetramer composed of a regulatory subunit (R) dimer and a catalytic subunit (C) dimer.
In this inactive state, the pseudosubstrate sequences on the R subunits block the active sites on the C subunits.
Three C subunit isoforms (C-alpha, C-beta, and C-gamma) and two families of regulatory subunits (RI and RII) with distinct cAMP binding properties have been identified.
The two R families exist in two isoforms, alpha and beta (RI-a, RI-b, RII-a, and RII-b).
Upon binding of cAMP to the R subunits, the autoinhibitory contact is eased and active monomeric C subunits are released.
PKA shares substrate specificity with Akt (PKB) and PKC, which are characterized by an arginine at position -3 relative to the phosphorylated serine or threonine residue (2).
Substrates that present this consensus sequence and have been shown to be phosphorylated by PKA are Bad (Ser155), CREB (Ser133), and GSK-3 (GSK-3a Ser21 and GSK-3b Ser9) (3-5).
In addition, combined knock-down of PKA C-a and -b blocks cAMP-mediated phosphorylation of Raf (Ser43 and Ser259) (6).
Autophosphorylation and phosphorylation by PDK-1 are two known mechanisms responsible for phosphorylation of the C subunit at Thr197 (7).
Applications:Suitable for use in Immunofluorescence and Western Blot.
Other applications not tested.
Recommended Dilution:Western Blot: 1:500-1:2000Immunofluorescence: 1:50-1:200Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20°C.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：200ul
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by affinity chromatography.
Form：Supplied as a liquid in PBS, pH 7.3, 0.02% sodium azide, 50% glycerol.
Specificity：Recognizes PRKAR1A. Species Crossreactivity: human, mouse, rat
Isotype：IgG
Calc Applications Abbrev：IF WB
Calc Crossreactivity：Hu Mo Rt
Immunogen：Recombinant protein corresponding to human PRKAR1A.
