预期应用
ELISA法定量测定大鼠血清、血浆、细胞培养物上清或其它相关液体中雌激素受体(ER)含量。
实验原理
本试剂盒应用双抗体夹心酶标免疫分析法测定标本中雌激素受体水平。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雌激素受体、生物素化的抗大鼠雌激素受体抗体、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雌激素受体呈正相关。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 total Estrogen Receptor,ER concentrations in cell culture supernates, serum, and plasma.
Introduction
The estrogen receptor (ER) is a member of the nuclear hormone family of intracellular receptors which is activated by the hormone 17β-estradiol. The main function of the estrogen receptor is as a DNA binding transcription factor which regulates gene expression. However the estrogen receptor also has additional functions independent of DNA binding. There are two different forms of the estrogen receptor usually referred to as α and β each encoded by a separate gene (ESR1 and ESR2 respectively). Hormone activated estrogen receptors form dimers, and since the two forms are coexpressed in many cell types, the receptors may form ERα (αα) or ERβ (ββ) homodimers or ERαβ (αβ) heterodimers.Estrogen receptor alpha and beta show significant overall sequence homology, and both are composed of seven domains.
Both ERs are widely expressed in different tissue types, however there are some notable differences in their expression patterns:The ERα is found in endometrium, breast cancer cells, ovarian stroma cells and in the hypothalamus.The expression of the ERβ protein has been documented in kidney, brain, bone, heart,lungs, intestinal mucosa, prostate, and endothelial cells.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for ER has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ER present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for ER 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 ER bound in the initial step. The color development is stopped and the intensity of the color is measured.
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