PKA RI-α/β Antibody

3927:

Immunofluorescence , Western Blotting

PKA RI-α/β Antibody detects endogenous levels of total PKA RI-α and PKA RI-β protein.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asn310 of human PKA RI-α. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from U118MG and CAD cells using PKA RI-α/β Antibody.

IF-IC

Confocal immunofluorescent analysis of U-118 MG cells using PKA RI-α/β Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

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-α, C-β, and C-γ) 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, α and β (RI-α, RI-β, RII-α, and RII-β). 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-3α Ser21 and GSK-3β Ser9) (3-5). In addition, combined knock-down of PKA C-α and -β 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).

1. Montminy, M. (1997) Annu. Rev. Biochem. 66, 807-822.
2. Dell'Acqua, M.L. and Scott, J.D. (1997) J. Biol. Chem. 272, 12881-12884.
3. Tan, Y. et al. (2000) J. Biol. Chem. 275, 25865-25869.
4. Gonzalez, G.A. and Montminy, M.R. (1989) Cell 59, 675-680.
5. Fang, X. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 11960-11965.
6. Dumaz, N. and Marais, R. (2003) J. Biol. Chem. 278, 29819 -29823.
7. Moore, M.J. et al. (2002) J. Biol. Chem. 277, 47878-47884.

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know !

• 4781 Phospho-PKA C (Thr197) Antibody
• 4782 PKA C-α Antibody
• 9621 Phospho-(Ser/Thr) PKA Substrate Antibody
• 9197 CREB (48H2) Rabbit mAb
• 9198 Phospho-CREB (Ser133) (87G3) Rabbit mAb
• 9323 Phospho-GSK-3β (Ser9) (5B3) Rabbit mAb
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7075 Anti-biotin, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
• 9998 BSA
• 9999 Nonfat Dry Milk

For Research Use Only. Not For Use In Diagnostic Procedures.