Cell Signaling Technology

Product Pathways - Ca / cAMP / Lipid Signaling

Phospho-PKA C (Thr197) (D45D3) Rabbit mAb #5661

No. Size Price
5661S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
5661T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
5661 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 42 Rabbit IgG

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting,

Specificity / Sensitivity

Phospho-PKA C (Thr197) (D45D3) Rabbit mAb detects endogenous levels of PKA C (-α, -β, and -γ) only when phosphorylated at Thr197.

Phospho-PKA C (Thr197) (D45D3)兔单克隆抗体可识别内源性的Thr197磷酸化的PKA C(-α, -β和-γ)。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr197 of human PKA C protein.

该单克隆抗体用与人类PKA C蛋白中Thr197位点附近的氨基酸序列对应的人工合成肽段免疫动物而制成。

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or λ phosphatase-treated, using Phospho-PKA C (Thr197) (D45D3) Rabbit mAb (upper) or PKA C-α Antibody #4782 (lower).

对NIH/3T3细胞抽提液,未处理或λ磷酸酶处理,使用Phospho-PKA C (Thr197) (D45D3)兔单抗(上图)或PKA C-抗体#4782(下图)进行Western blot分析。

Background

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).

第二信使环磷酸腺苷(cAMP)在哺乳动物细胞中激活cAMP依赖的蛋白激酶(PKA或者cAPK),控制如基因转录,离子转运和蛋白磷酸化等许多细胞内机制(1)。失活的PKA的是一个由调节亚基(R)和催化亚基(C)组成的异二聚体。在失活状态时,R亚基的假底物序列阻止了C亚基的活性中心。已识别出三个催化亚基的亚型(C-α, C-β, 和C-γ)和两个调节亚基家族(RI和RII),其分别具有不同的cAMP结合特性。两个R家族又进一步存在两种亚型,α和β (RI-α, RI-β, RII-α和RII-β)。R亚基与cAMP结合后,自身抑制性接触被解除,有活性的C亚基单体被释放。PKA与Akt(PKB)和PKC共有相同的底物特异性,以一个在3位的与酪氨酸和苏氨酸残疾磷酸化相关的精氨酸为特点(2)。具有这些相同结构序列的底物已经被证实能被PKA磷酸化,它们包括Bad (Ser155), CREB (Ser133), 和GSK-3 (GSK-3α Ser21和GSK-3β Ser9)(3-5)。另外,同时下调PKA C-α和-β可以阻止cAMP介导的Raf(Ser43和Ser259)磷酸化(6)。自身磷酸化和被PDK-1磷酸化是C亚基Thr197磷酸化的两个已知的机制(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.

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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