PhosphoPlus? IκBα (Ser32/36) Antibody Kit

产品编号：CST-9240S 品牌：CST 原厂货号：9240S
产品分类：生化和细胞分析 > 细胞分析试剂盒 > 细胞通路检测试剂盒
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包装：

运保温度:

–20°C

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其它组分：

IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) 【子货号：#4814，包装：100 µl，，运保温度：–20°C】

20X LumiGLO® Reagent and 20X Peroxide 【子货号：#7003，包装：5 ml each，，运保温度：–20°C】

Anti-biotin, HRP-linked Antibody 【子货号：#7075，包装：100 µl，，运保温度：–20°C】

Anti-mouse IgG, HRP-linked Antibody 【子货号：#7076，包装：100 µl，，运保温度：–20°C】

Biotinylated Protein Ladder Detection Pack 【子货号：#7727，包装：100 µl，，运保温度：–20°C】

NF-κB Control Cell Extracts【子货号： #9243，包装：200 µl，，运保温度：–20°C】

Phospho-IκBα (Ser32/36) (5A5) Mouse mAb 【子货号：#9246，包装：100 µl，，运保温度：–20°C】

描述：

IκBα (L35A5) Mouse mAb (氨基端抗原) #4814 能够检测内源水平IκBα总蛋白水平。Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246 能够检测内源性在Ser32/36位点发生磷酸化的IκBα蛋白。这两个抗体均不与磷酸化的IκB家族的其他蛋白交叉反应。IκBα (L35A5) Mouse mAb (氨基端抗原) #4814是通过合成对应人源IκBα N-末端肽段的GST-IκBα融合蛋白免疫小鼠而获得。Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246是通过合成人源性对应IκBα Ser32/36位点周围的磷酸化肽段来免疫小鼠而获得。未经处理的HeLa细胞的总细胞提取液作为阴性对照，保存在SDS样品缓冲液中。经TNF-α(#2169, 20 ng/ml,5 min)处理的HeLa细胞的总细胞提取液作为阳性对照，保存在SDS样品缓冲液中。PhosphoPlus® IκBα (Ser32/36) Antibody Kit 为在Ser32/36位点磷酸化的IκBα 的研究提供了试剂盒步骤指导。试剂盒中包含了总IκBα蛋白抗体，特异位点磷酸化的抗体，总细胞裂解液的阳性/阴性对照和Western免疫印迹实验所需的二抗和试剂。 NF-κB/Rel转录调控因子在细胞质中与IκB抑制蛋白结合以非活性形式存在(1-3)。磷酸化IκBα 的Ser32 和 Ser36位点后，蛋白水解酶体介导的降解使得 NF-κB释放激活并入核(3-7)。IκBα的磷酸化导致了Rel依赖的转录能够被很多胞外信号激活，包括炎症因子，生长因子和趋化因子。能够磷酸化IκB这些活性位点的激酶已经得到了鉴定(8)。

原厂资料：

Specificity / Sensitivity

IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814 detects endogenous levels of total IκBα protein. Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246 detects endogenous levels of IκBα only when phosphorylated at Ser32/36. Neither antibody cross-reacts with other IκB family members at physiological levels.

Source / Purification

IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814 is produced by immunizing mice with a GST-IκBα fusion protein corresponding the amino-terminus of human IκBα. Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246 is produced by immunizing mice with a synthetic phosphopeptide corresponding to residues surrounding Ser32/36 of human IκBα. Total cell extracts from HeLa cells prepared without treatment serve as a negative control. Supplied in SDS Sample Buffer. Total cell extracts from HeLa cells prepared with TNF-α treatment (#2169, 20 ng/ml for 5 minutes) serve as a positive control. Supplied in SDS Sample Buffer.

Description

The PhosphoPlus® IκBα (Ser32/36) Antibody Kit provides reagents and protocols for the rapid analysis of IκBα phosphorylation at Ser32/36. The kit contains a total IκBα antibody, a phospho-specific antibody, and positive/negative control whole cell lysates, along with secondary antibodies and reagents for Western blotting.

Background

The NF-κB/Rel transcription factors are present in the cytosol in an inactive state complexed with the inhibitory IκB proteins (1-3). Activation occurs via phosphorylation of IκBα at Ser32 and Ser36 followed by proteasome-mediated degradation that results in the release and nuclear translocation of active NF-κB (3-7). IκBα phosphorylation and resulting Rel-dependent transcription are activated by a highly diverse group of extracellular signals including inflammatory cytokines, growth factors, and chemokines. Kinases that phosphorylate IκB at these activating sites have been identified (8).

注意事项：

Storage: Antibodies are supplied in 10 mM sodium HEPES
(pH 7.5), 150 mM NaCI, 100 μg/ml BSA and 50% glycerol.
Store at –20°C. Do not aliquot the antibodies. NF-κB Control
Cell Extracts are supplied in SDS Sample Buffer: 62.5
mM Tris-HCl (pH 6.8 at 25°C), 2% w/v SDS, 10% glycerol,
50 mM DTT, 0.01% w/v bromophenol blue or phenol red.
Store at –20°C or below.