| 货号 | 7295C |
| 描述 | CSTs PathScan® Total HSP27 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total HSP27 protein. An HSP27 Antibody has been coated onto the microwells. After incubation with cell lysates, both nonphospho- and phospho-HSP27 are captured by the coated antibody. Following extensive washing, an HSP27 Mouse mAb is added to detect the captured HSP27 protein. HRP-linked anti-mouse antibody #7076* is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total HSP27 protein. * Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human/Monkey |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Total HSP27 Sandwich ELISA Kit detects endogenous levels of total HSP27 protein. Using PathScan® Phospho-HSP27 (Ser78) Sandwich ELISA Kit #7290, a significant induction of phospho-HSP27 (Ser78) in HeLa cells treated with UV light can be detected. However, the level of total HSP27 (phospho and non-phospho), detected by this Sandwich ELISA Kit #7295, remains unchanged (Figure 1). COS cells treated with UV light show similar results (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. |
| 供应商 | CST |
| 背景 | Heat shock protein (HSP) 27 is one of the small HSPs that are constitutively expressed at different levels in various cell types and tissues. Like other small heat shock proteins, HSP27 is regulated at both the transcriptional and posttranslational levels (1). In response to stress, the expression level of HSP27 increases several-fold to confer cellular resistance to the adverse environmental change. HSP27 is phosphorylated at Ser15, Ser78 and Ser82 by MAPKAP kinase 2 as a result of the activation of the p38 MAP kinase pathway (2,3). Phosphorylation of HSP27 causes a change in its tertiary structure, which shifts from large homotypic multimers to dimers and monomers (4). It has been shown that phosphorylation and increased concentration of HSP27 modulates actin polymerization and reorganization (5,6). |
| 存放说明 | 4C |
| 参考文献 | Arrigo, A.P. and Landry, J. (1994) Cold Spring Harbor Laboratory Press, NY, 335-373. Landry, J. et al. (1992) J. Biol. Chem. 267, 794-803. Rouse, J. et al. (1994) Cell 78, 1027-1037. Rogalla, T. et al. (1999) J. Biol. Chem. 274, 18947-18956. Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214. Rousseau, S. et al. (1997) Oncogene 15, 2169-2177. |
Figure 1: Treatment of HeLa cells with UV light stimulates phosphorylation of HSP27 at Ser78, detected by PathScan® Phospho-HSP27 (Ser78) Sandwich ELISA kit, #7290, but does not affect the level of total HSP27 protein detected by PathScan® Total HSP27 Sandwich ELISA kit, #7295. OD450 readings (upper) and the corresponding Western blot using Phospho-HSP27 (Ser78) Ab #2405 (lower right) or HSP27 (G31) Mouse mAb #2402 (lower left), are shown. 图1：紫外光处理后的HeLa细胞产生了HSP27的Ser78磷酸化，为PathScan Phospho-HSP27 (Ser78) Sandwich ELISA kit #7290所检测到，但PathScan Total HSP27 Sandwich ELISA kit #7295可检测到的总HSP27水平的确没有变化。上图为OD450结果，右下为使用Phospho-HSP27 (Ser78) Ab #2405进行的Western blot，左下为使用HSP27 (G31) Mouse mAb #2402进行的Western blot。 | |
Figure 2: Relationship between protein concentration of lysates from untreated and UV-treated HeLa cells and kit assay optical density readings. HeLa cells (70-85% confluence) were treated with UV and lysed after incubation at 37oC for 30 minutes.图2：显示未处理和紫外线处理的HeLa细胞裂解液中的蛋白浓度与试剂盒光学浓度读数的关系。HeLa细胞（70-85%融合）在37℃下孵育30分钟后用紫外线处理裂解。 |
