PathScan® Total HSP27 Sandwich ELISA Kit (#7295) Datasheet with Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes	Product #	Quantity	Color	Storage Temp
HSP27 Rabbit mAb Coated Microwells	42806	96 tests		+4C
HSP27 Mouse Detection mAb	14260	1 ea	Green (Lyophilized)	+4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)	13304	1 ea	Red (Lyophilized)	+4C
Detection Antibody Diluent	13339	11 ml	Green	+4C
HRP Diluent	13515	11 ml	Red	+4C
TMB Substrate	7004	11 ml		+4C
STOP Solution	7002	11 ml		+4C
Sealing Tape	54503	2 ea		+4C
ELISA Wash Buffer (20X)	9801	25 ml		+4C
ELISA Sample Diluent	11083	25 ml	Blue	+4C
Cell Lysis Buffer (10X)	9803	15 ml		-20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

CST's 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. Anti-mouse IgG, HRP-linked antibody 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.

Specificity/Sensitivity

CST's 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.

Background

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 HSPs, HSP27 is regulated at both the transcriptional and posttranslational levels (1). In response to stress, the HSP27 expression increases several-fold to confer cellular resistance to the adverse environmental change. HSP27 is phosphorylated at Ser15, Ser78, and Ser82 by MAPKAPK-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).

Background References

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

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PathScan is a registered trademark of Cell Signaling Technology, Inc.

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Limited Uses

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