本试剂盒应用双抗体夹心酶标免疫分析法测定标本中HSF1水平。用纯化的抗体包被微孔板，制成固相抗体，往包被抗体的微孔中依次加入HSF1、生物素化的抗大鼠HSF1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HSF1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 rat Heat Shock Factor 1,HSF1 concentrations in cell culture supernates, serum, and plasma.
Introduction
Heat shock proteins (HSP) are a group of proteins whose expression is increased when the cells are exposed to elevated temperatures or other stress. This increase in expression is transcriptionally regulated. This dramatic upregulation of the heat shock proteins induced mostly by Heat Shock Factor (HSF) is a key part of the heat shock response.The function of heat-shock proteins is similar in virtually all living organisms, from bacteria to humans.
It was discovered that Heat Shock Factor 1(HSF1) is a powerful multifaceted modifier of carcinogenesis. HSF1 knockout mice show significantly decreased incidence of skin tumor after topical application of DMBA, a mutagen .HSF1 is potent and activated rapidly though transiently by heat shock, whereas HSF2 is a less active transcriptional regulator but can retain its DNA binding properties for extended periods. In vertebrates, HSF1 mediates the ubiquitous response to stress stimuli, while the role of a second HSE-binding factor, HSF2, is still unclear. HSF2 shows common structural domains with HSF1; however, it is active at normal temperatures.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for HSF1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any HSF1 present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for HSF1 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 HSF1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
