| 货号 | 4782S |
| 反应种属 | Human/Mouse/Rat |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/F |
| 目标/特异性 | PKA C-α Antibody detects endogenous levels of total PKA C-α. |
| 使用方法 | WB(1:1000) IP (1:50) F (1:50) IF-IC (1:100) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | 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). |
| 存放说明 | -20C |
| 计算分子量 | 42 |
| 参考文献 | 1 . Montminy, M. (1997) Annu Rev Biochem 66, 807-22. 2 . DellAcqua, M.L. and Scott, J.D. (1997) J Biol Chem 272, 12881-4. 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, 29819–23 7 . Moore, M.J. et al. (2002) J. Biol. Chem. 277, 47878-47884. |
Western blot analysis of extracts from HeLa, C6, PC12 and NIH/3T3 cells, using PKA C-α Antibody.对HeLa、C6、PC12和NIH/3T3细胞，使用PKA C-α抗体进行Western blot分析。 | |
Flow cytometric analysis of HeLa cells, using PKA C-α Antibody (blue) compared to a nonspecific negative control antibody (red).使用PKA C-α抗体（蓝）和非特异的阴性对照抗体（红），对HeLa细胞进行流式细胞仪分析。 | |
Confocal immunofluorescent analysis of HeLa cells using PKA C-alpha Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).对HeLa细胞使用PKA C-α抗体（绿色）进行共聚焦免疫荧光分析。肌动蛋白用DY-554 phalloidin（红色）标记。蓝色伪彩=DRAQ5?(荧光DNA染料)。 | |
Western blot analysis of extracts from HeLa cells transfected with non-targeted (-) or PKA C-α (+) siRNA. PKA C-α was detected using the PKA C-α Antibody #4782, and Akt1 was detected using Akt1 (2H10) Monoclonal Antibody #2967. The PKA C-α Antibody confirms silencing of PKA C-α expression, and the Akt1 Antibody is used to control for loading and specificity of PKA C-α siRNA.对空白转染（-）或PKA C-α（+）siRNA的HeLa细胞进行Western blot分析。PKA C-α使用PKA C-α抗体#4782检测，Akt1使用Akt1（2H10）单克隆抗体#2967检测。PKA C-α抗体证实PKA C-α的表达被沉默，Akt1抗体用作显示PKA C-α siRNA装载和特异性的对照。 | |
Western blot analysis of extracts from HeLa cells, transfected with either 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-), SignalSilence® PKA C-α siRNA II (+) or SignalSilence® PKA C-α siRNA I #6406 (+), using PKA C-α Antibody #4782 and α-Tubulin (11H10) Rabbit mAb #2125. The PKA C-α antibody confirms silencing of PKA C-α expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of PKA C-α siRNA.对转染100 nM SignalSilence? Control siRNA (Fluorescein Conjugate) #6201 (-)、 SignalSilence? PKA C-α siRNA II(+) 或者SignalSilence? PKA C-α siRNA I #6406 (+)的HeLa细胞使用PKA C-α抗体#4782和α-Tubulin (11H10)兔单克隆抗体#2125进行Western blot分析。PKA C-α抗体证实PKA C-α的表达被沉默，α-Tubulin (11H10)兔单克隆抗体用作显示PKA C-α siRNA装载和特异性的对照。 |
