Phospho-Insulin/IGF Receptor Antibody Sampler Kit

• 产品编号：CST-8338S      品牌：Cell Signaling Technology       原厂货号：8338S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
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描述：

Phospho-Insulin/IGF Receptor Antibody Sampler Kit中的每种抗体都识别内源性特异性的靶蛋白水平，活化态特异性的抗体只识别在特定位点磷酸化的靶蛋白。IGF-I Receptor beta (111A9) Rabbit mAb用于检测内源性IGF-I beta受体总蛋白水平，不与胰岛素受体交叉反应。Phospho-IGF-I Receptor beta (Tyr1135) (DA7A8) Rabbit mAb用于检测在Tyr1135位点磷酸化的内源性IGF-I受体，该抗体可与胰岛素受体的Tyr1150交叉反应，同样也可能与其它一些过表达的酪氨酸磷酸化的酪氨酸激酶交互反应。Phospho-IGF-I Receptor β (Tyr1131)/Insulin Receptor β (Tyr1146) Antibody用于检测内源性Tyr1131位点磷酸化的IGF-I受体和Tyr1146磷酸化的胰岛素受体，该抗体可与激活的PDGF, FGF和EGF受体，以及ErbB2和c-Met交互反应。Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb用于检测内源性在酪氨酸1135/1136或1150/1151位点磷酸化的IGF-I和胰岛素受体，并且该抗体不与其它的酪氨酸磷酸化的酪氨酸激酶交互反应。Phospho-IGF-I Receptor β (Tyr980) (C14A11) Rabbit mAb用于检测内源性在Tyr980位点磷酸化的IGF-I受体β，并且不与激活型的胰岛素受体和FLT3交互反应。Insulin Receptor β (4B8) Rabbit mAb用于检测内源性总的胰岛素受体β，并且不与IGF-I受体β交互反应。IGF-I受体β、人源IGF-I受体βTyr1135/1136位点、人源IGF-I受体βTyr980位点等活化态特异性的多抗通过用与人源IGF-I受体β蛋白相应位点周围残基序列相同的合成肽免疫动物制备而得。抗体的纯化采用蛋白A与肽段亲和层析相结合的方法。Phospho-Insulin/IGF Receptor Antibody Sampler Kit为检测内源性胰岛素受体和IGF-I受体β蛋白的总蛋白水平，以及在特定位点磷酸化的两种受体的水平，提供一套经济有效的解决方案。试剂盒中的每种抗体都至少能够满足四次Western blot实验所需的一抗。一型胰岛素样生长因子受体(IGF-IR)是一种跨膜的酪氨酸激酶，在许多新生及成熟的组织细胞中都广泛表达(1-3)。IGF-I和IGF-II配体结合到受体上去，受体就发生自我磷酸化。在激酶结构域，有三个酪氨酸残基(Tyr1131, Tyr1135,Tyr1136)是最先发生自身磷酸化的位点(4)。这三个残基的磷酸化对激酶的激活是必需的(5,6)。胰岛素受体(IRs)跟胰岛素样生长因子受体有着相似的结构和功能，在它的激酶活化环上同样有三个酪氨酸簇(Tyr1146/1150/1151)。胰岛素受体酪氨酸残基的磷酸化是细胞接受胰岛素刺激后最先发生的生化反应(7)。自身磷酸化最先在Tyr1146和Tyr1150(或Tyr1151)发生，但激酶的激活需要三者同时磷酸化(8)。

1. Adams, T.E. et al. (2000) Cell Mol Life Sci 57, 1050-93.
2. Baserga, R. (2000) Oncogene 19, 5574-81.
3. Scheidegger, K.J. et al. (2000) J Biol Chem 275, 38921-8.
4. Hernández-Sánchez, C. et al. (1995) J Biol Chem 270, 29176-81.
5. Lopaczynski, W. et al. (2000) Biochem Biophys Res Commun 279, 955-60.
6. Baserga, R. (1999) Exp Cell Res 253, 1-6.
7. White, M.F. et al. (1985) J Biol Chem 260, 9470-8.
8. White, M.F. et al. (1988) J Biol Chem 263, 2969-80.

原厂资料：

Specificity / Sensitivity

Each antibody in the Phospho-Insulin/IGF Receptor Antibody Sampler Kit recognizes endogenous levels of the specific target protein, with activation state-specific antibodies recognizing target proteins only when phosphorylated at the indicated residues. 
 IGF-I Receptor beta (111A9) Rabbit mAb detects endogenous levels of total IGF-I receptor beta protein and does not cross-react with insulin receptor. 
 Phospho-IGF-I Receptor beta (Tyr1135) (DA7A8) Rabbit mAb detects endogenous levels of IGF-I receptor only when phosphorylated at Tyr1135. This antibody cross-reacts with Tyr1150 of insulin receptor and may also cross-react with other overexpressed related tyrosine-phosphorylated tyrosine kinases. 
 Phospho-IGF-I Receptor β (Tyr1131)/Insulin Receptor β (Tyr1146) Antibody detects endogenous levels of Tyr1131-phosphorylated IGF-I receptor and Tyr1146-phosphorylated insulin receptor. This antibody cross-reacts with activated PDGF, FGF and EGF receptors; ErbB2; and c-Met. 
 Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb detects endogenous levels of IGF-I receptor and insulin receptor only when phosphorylated at tyrosine 1135/1136 or tyrosine 1150/1151, and does not cross-react with other related tyrosine-phosphorylated tyrosine kinases. 
 Phospho-IGF-I Receptor β (Tyr980) (C14A11) Rabbit mAb detects endogenous levels of IGF-I receptor β protein only when phosphorylated at Tyr980, and may cross-react with activated insulin receptors and FLT3. 
 Insulin Receptor β (4B8) Rabbit mAb detects endogenous levels of total insulin receptor β and does not cross-react with IGF-I receptor β.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy-terminal residues of human IGF-IR β or residues surrounding Tyr960 of human insulin receptor β. Activation state-specific monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1135 of human IGF-I Receptor β, Tyr1135/1136 of human IGF-I Receptor β, or Tyr980 of human IGF-I Receptor β. Activation state-specific polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues of human IGF-I Receptor β. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Description

The Phospho-Insulin/IGF Receptor Antibody Sampler Kit provides an economical means of evaluating total Insulin Receptor and IGF-I Receptor β protein levels as well as Insulin and IGF-I Receptor β phosphorylated at specific sites. The kit includes enough antibody to perform four western blot experiments with each primary antibody.

Background

Type I insulin-like growth factor receptor (IGF-IR) is a transmembrane receptor tyrosine kinase that is widely expressed in many cell lines and cell types within fetal and postnatal tissues (1-3). Receptor autophosphorylation follows binding of the IGF-I and IGF-II ligands. Three tyrosine residues within the kinase domain (Tyr1131, Tyr1135, and Tyr1136) are the earliest major autophosphorylation sites (4). Phosphorylation of these three tyrosine residues is necessary for kinase activation (5,6). Insulin receptors (IRs) share significant structural and functional similarity with IGF-I receptors, including the presence of an equivalent tyrosine cluster (Tyr1146/1150/1151) within the kinase domain activation loop. Tyrosine autophosphorylation of IRs is one of the earliest cellular responses to insulin stimulation (7). Autophosphorylation begins with phosphorylation of Tyr1146 and either Tyr1150 or Tyr1151, while full kinase activation requires triple tyrosine phosphorylation (8).

1. Adams, T.E. et al. (2000) Cell Mol Life Sci 57, 1050-93.
2. Baserga, R. (2000) Oncogene 19, 5574-81.
3. Scheidegger, K.J. et al. (2000) J Biol Chem 275, 38921-8.
4. Hernández-Sánchez, C. et al. (1995) J Biol Chem 270, 29176-81.
5. Lopaczynski, W. et al. (2000) Biochem Biophys Res Commun 279, 955-60.
6. Baserga, R. (1999) Exp Cell Res 253, 1-6.
7. White, M.F. et al. (1985) J Biol Chem 260, 9470-8.
8. White, M.F. et al. (1988) J Biol Chem 263, 2969-80.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibodies.