Phospho-PKC (pan) (gamma Thr514) Antibody

• 产品编号：CST-9379S      品牌：CST       原厂货号：9379S
• 产品分类：抗体 > 一抗 > 磷酸化抗体
• 应用分类：

描述：

Phospho-PKC (pan) (γ Thr514)抗体可识别内源性的与人类PKCγThr514磷酸化同源的磷酸化的PKC α、βI、βII、γ、δ、ε、η、θ亚型。此抗体不与其它位点磷酸化的PKC蛋白发生交叉反应。该多克隆抗体用与人类PKCγ中Thr514位点附近的氨基酸序列对应的人工合成磷酸化的肽段免疫动物而制备。该抗体使用蛋白A和蛋白亲和层析纯化而得。蛋白激酶C（PKC）的激活是最早的级联事件之一，控制各种细胞反应，包括分泌、基因表达、细胞增殖和肌肉收缩（1,2）。PKC亚型属于基于钙依赖性和活化的三个群体。经典PKCs都是钙依赖性的，通过它们的C2结构域，由磷脂酰丝氨酸(PS)、甘油二酯(DAG)和佛波酯(TPA, PMA)通过其富含半胱氨酸的C1结构域激活。新型和非经典型PKCs都是非钙依赖性的，但只有新型PKCs可以被PS、DAG和佛波酯激活(3-5)。这三种PKC群的成员包含一个假底物或自抑制结构域，可以与催化区域的底物结合位点结合，以防止在缺少辅酶因子或激活剂的情况下激活。PKC活性的控制通过三个独特的磷酸化事件调节。在体内，通过自身磷酸化，磷酸化活化环中的苏氨酸500位点，641位点，和羧基末端的疏水性丝氨酸660位点（2）。非经典型PKC缺乏疏水性区域的磷酸化，与谷氨酸而不是在更多典型PKC亚型中发现的丝氨酸或苏氨酸残基有关。 PDK1或相关酶负责PKC的激活。最近PKCμ（PKD）成为PKC超家族新的一员，通过其C1结构域受DAG和TPA调节。PKD的特点是有一个PH结构域，并有其独特的底物识别和高尔基体定位（6）。PKC相关激酶（PRK）缺少C1域，不响应DAG或佛波醇酯。磷脂脂质激活PRKs，小Rho-家族GTP酶结合同源性区域1（HR1），以调节PRK激酶活性（7）。

1. Nishizuka, Y. (1984) Nature 308, 693-8.
2. Keranen, L.M. et al. (1995) Curr Biol 5, 1394-1403.
3. Mellor, H. and Parker, P.J. (1998) Biochem J 332 ( Pt 2), 281-92.
4. Ron, D. and Kazanietz, M.G. (1999) FASEB J 13, 1658-76.
5. Moscat, J. and Diaz-Meco, M.T. (2000) EMBO Rep 1, 399-403.
6. Baron, C.L. and Malhotra, V. (2002) Science 295, 325-8.
7. Flynn, P. et al. (2000) J Biol Chem 275, 11064-70.

原厂资料：

Specificity / Sensitivity

Phospho-PKC (pan) (gamma Thr514) Antibody detects endogenous levels of PKC alpha, beta I, beta II, gamma, delta, epsilon, eta and theta isoforms only when phosphorylated at a residue homologous to threonine 514 of human PKCgamma. This antibody does not detect PKC phosphorylated at other sites.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr514 of human PKCgamma. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Activation of protein kinase C (PKC) is one of the earliest events in a cascade that controls a variety of cellular responses, including secretion, gene expression, proliferation, and muscle contraction (1,2). PKC isoforms belong to three groups based on calcium dependency and activators. Classical PKCs are calcium-dependent via their C2 domains and are activated by phosphatidylserine (PS), diacylglycerol (DAG), and phorbol esters (TPA, PMA) through their cysteine-rich C1 domains. Both novel and atypical PKCs are calcium-independent, but only novel PKCs are activated by PS, DAG, and phorbol esters (3-5). Members of these three PKC groups contain a pseudo-substrate or autoinhibitory domain that binds to substrate-binding sites in the catalytic domain to prevent activation in the absence of cofactors or activators. Control of PKC activity is regulated through three distinct phosphorylation events. Phosphorylation occurs in vivo at Thr500 in the activation loop, at Thr641 through autophosphorylation, and at the carboxy-terminal hydrophobic site Ser660 (2). Atypical PKC isoforms lack hydrophobic region phosphorylation, which correlates with the presence of glutamic acid rather than the serine or threonine residues found in more typical PKC isoforms. The enzyme PDK1 or a close relative is responsible for PKC activation. A recent addition to the PKC superfamily is PKCμ (PKD), which is regulated by DAG and TPA through its C1 domain. PKD is distinguished by the presence of a PH domain and by its unique substrate recognition and Golgi localization (6). PKC-related kinases (PRK) lack the C1 domain and do not respond to DAG or phorbol esters. Phosphatidylinositol lipids activate PRKs, and small Rho-family GTPases bind to the homology region 1 (HR1) to regulate PRK kinase activity (7).

1. Nishizuka, Y. (1984) Nature 308, 693-8.
2. Keranen, L.M. et al. (1995) Curr Biol 5, 1394-1403.
3. Mellor, H. and Parker, P.J. (1998) Biochem J 332 ( Pt 2), 281-92.
4. Ron, D. and Kazanietz, M.G. (1999) FASEB J 13, 1658-76.
5. Moscat, J. and Diaz-Meco, M.T. (2000) EMBO Rep 1, 399-403.
6. Baron, C.L. and Malhotra, V. (2002) Science 295, 325-8.
7. Flynn, P. et al. (2000) J Biol Chem 275, 11064-70.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 μg/ml BSA and 50% glycerol. Store at –20°C.
Do not aliquot the antibody.