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重要规格表
检测方法 Colorimetric 描述 概述 Detects and quantifies the level of phosphorylated Hsp27 protein in cell lysates, tissue extracts, and biological fluids. Hsp27 is the small heat shock protein that acts as a molecular chaperone interacting with a large number of proteins. Hsp27 is known for its ability to negatively regulate the mitochondrial pathway to caspase activation. The phosphorylated form of Hsp27 also plays a major role in apoptosis through its ablity to prevent cell death by maintaining redox homeostasis and mitochondrial stability. 产品目录编号 CBA078 品牌系列 Calbiochem®
应用数据 
A standard curve was generated using phosphorylated (black square) and unphosphorylated (black triangle) Hsp27 as outlined in the Detailed Protocol. Assay Range: 0.1-10 ng/ml; lower limit of detection: ~0.1 ng/ml.
Various tumor cell lines (indicated in the figure) were exposed to 100 J/m2 UV irradiation or treated with 400 mM sorbitol for 30 min. Cell lysates were prepared using CytoBuster™ Protein Extraction Reagent (Cat. No. 71009) according to the recommended protocol. The level of phosphorylated Hsp27 was determined as outlined in the Detailed Protocol and expressed as ng phosphorylated Hsp27/mg total protein.
(A) MCF-7 and HeLa cells were either left untreated (-UV) or exposed to UV-C radiation (+UV). Cell lysates were prepared using CytoBuster™ Protein Extraction Reagent (Cat. No. 71009), subjected to SDS-PAGE, and transferred to nitrocellulose membrane for Western blotting analysis using Anti-Phospho-Hsp27 Detector Antibody Lane 1: MWM; lane 2: MCF-7, -UV; lane 3: MCF-7, +UV; lane 4: HeLa, -UV; lane 5: HeLa, +UV. (B) Unphosphorylated Hsp27 and recombinant Hsp27 that had been phosphorylated in vitro with MAPKAP-2 kinase were subjected to SDS-PAGE and Western blotting analysis using Anti-Phospho-Hsp27 Detector Antibody. Lane 1: MWM; lane 2: in vitro phosphorylated, recombinant Hsp27; lane 3: unphosphorylated, recombinant Hsp27.
MCF7 cells were either left untreated (left bar) or treated with a highly specific, cell-permeable inhibitor of p38 MAP kinase, SB203580 (Cat. No. 559389), (2 µM) followed by UV treatment (right bar). Cell lysates were prepared using CytoBuster™ Protein Extraction Reagent (Cat. No. 71009) and the level of phosphorylated Hsp27 was determined as outlined in the Detailed Protocol.
参考 参考 Xu, L., et al. 2006. Oncogene 25, 2987.
Shin, K.D., et al. 2005. J. Biol. Chem. 280, 41439.
Whitesell, L. and Lindquist, S. I. 2005. Nature Rev. 5, 761.
Eustace, B.K., et al. 2004. Nature Cell Biol. 6, 507.
Bruey, J-M., et al. 2000. Nature Cell Biol. 2, 645.
Cornford, P.A., et al. 2000. Cancer Res. 60, 7099.
Garrido, C., et al. 1999. FASEB J. 13, 2061.产品信息 检测方法 Colorimetric 形式 96 Tests 格式 96-well plate 试剂盒包含 Hsp27-Coated Plate, Rabbit Anti-Hsp27 Detector Antibody, Phosphorylated Hsp27 Standard, HRP-Conjugate, Assay Diluent, ELISA 20X Plate Wash Concentrate, TMB Substrate, ELISA Stop Solution, and a user protocol 正控制 Phosphorylated Hsp27 Standard, UV-treated HeLa or MCF7 cells or sorbitol-treated DU145 cells 生物信息 化验范围 0.1-10 ng/ml 化验时间 4.5 h 样品类型 Cell lysates 产品使用声明 预定用途 The Calbiochem® PhosphoDetect™ Hsp27 (pSer78/82) ELISA Kit is intended for the quantitative measurement of Hsp27 that is phosphorylated at Ser⁷⁸ and Ser⁸². The assay has been validated with human cell lysates and does not detect unphosphorylated Hsp27. 储存和货运信息 货运代码 Blue Ice Only
毒性 Regulatory Review
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储存 -20°C
存储条件 Upon arrival store the entire contents of the kit at -20°C. 无需冷冻 No
补充信息 试剂盒包含 Hsp27-Coated Plate, Rabbit Anti-Hsp27 Detector Antibody, Phosphorylated Hsp27 Standard, HRP-Conjugate, Assay Diluent, ELISA 20X Plate Wash Concentrate, TMB Substrate, ELISA Stop Solution, and a user protocol
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