本试剂盒应用双抗体夹心酶标免疫分析法测定标本中HSP27水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入HSP27抗原、生物素化的抗人HSP27抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的HSP27呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。
Intended use
This immunoassay kit allows for the specific measurement of human HSP27 concentrations in cell culture supernates, serum, and plasma.
Introduction
Heat shock protein (HSP) 27 is one of the small HSPs. 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).
1.Arrigo, A.P. and Landry, J. (1994) Cold Spring Harbor Laboratory Press, NY, 335-373.
2.Landry, J. et al. (1992) J. Biol. Chem. 267, 794-803.
3.Rouse, J. et al. (1994) Cell 78, 1027-1037.
4.Rogalla, T. et al. (1999) J. Biol. Chem. 274, 18947-18956.
5.Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
6.Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for HSP27 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any HSP27 present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for HSP27 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells
and color develops in proportion to the amount of HSP27 bound in the initial step. The color
development is stopped and the intensity of the color is measured.
