PathScan® Total IκBα Sandwich ELISA Kit【科瑞奥博】

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PathScan® Total IκBα Sandwich ELISA Kit

货号： 7360C 基本售价： 6346.0 元规格： 1Kit

产品信息

概述
货号	7360C
反应种属	Human/Mouse
应用	ELISA
目标/特异性	CSTs PathScan® Total IkappaB-alpha Sandwich ELISA Kit detects endogenous levels of total IκBα protein. Using PathScan® Phospho-

性能
供应商	CST
背景	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).
存放说明	4C
参考文献	Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6. Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70. Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8. Brown, K. et al. (1995) Science 267, 1485-8. Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83. Chen, Z.J. et al. (1996) Cell 84, 853-62. Karin, M. and Ben-Neriah, Y. (2000) Annu Rev Immunol 18, 621-63. Chen, J. et al. (2008) J Cell Mol Med , .

参考图片

Figure 1: Treatment of HeLa cells with TNF-α stimulates phopsphorylation of IκBα at Ser32, detected by PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit, #7355, but does not affect the level of total IκBα protein detected by PathScan® Total IκBα Sandwich ELISA Kit, #7360. Treatment with MG132, a proteasome inhibitor, (37ºC for 180 min before TNF-α induction) causes accumulation of phospho-IκBα in control and TNF-α-treated cells, shown in both Sandwich ELISA and Western blot analysis. OD 450 readings are shown in the top figure, while the corresponding Western blot using Phospho-IκBα (Ser32) Ab #9241 (right panel) or IκBα (L27H11) Mouse mAb #7361 (left panel), is shown in the bottom figure.

用TNF-α处理HeLa3细胞后使IκBα蛋白丝氨酸（32位点）磷酸化，检测试剂盒是PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit, #7355，但是总IκBα蛋白水平并没有受到影响，检测试剂盒是PathScan® Total IκBα Sandwich ELISA Kit, #7360。Sandwich ELISA 和Western blot检测分析都显示用MG132，一个蛋白酶体抑制子处理对照细胞裂解物（在用TNF-α诱导之前37ºC，180 min）后，导致对照组和TNF-α-处理的细胞中phospho-IκBα的堆积。上图显示OD450读数，下图是对应的Western blot检测结果，使用的抗体是Phospho-IκBα (Ser32) Ab #9241 (右侧) 和 IκBα (L27H11) Mouse mAb #7361 (左侧)。

Figure 2: Linear relationship between protein concentration of lysates from untreated and TNF-α-treated HeLa cells and kit assay optical density readings. HeLa cells (70-85% confluence) were treated with TNFα (10 ng/ml) and lysed after incubation at 37ºC for 5 minutes.

图2：未处理、TNF-α处理的HeLa细胞裂解物中蛋白质浓度和试剂盒分析光学密度读数之间的线性关系。TNFα (10 ng/ml)处理的HeLa细胞（70-85%生长汇合）37ºC孵育5min后再裂解。