预期应用
ELISA法定量测定大鼠血清、血浆或其它相关液体中TGF-β诱导早期基因1(TIEG1)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TIEG1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TIEG1抗原、生物素化的抗大鼠TIEG1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TIEG1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 TGF-beta-inducible early response gene-1,TIEG1 concentrations in cell culture supernates, serum and plasma.
Introduction
ignaling through the Transforming Growth Factor- (TGF)/Smad pathway plays an important role in a variety of cellular processes including the control of cellular proliferation and oncogenesis. The TGF Inducible Early Gene (TIEG) is a primary response gene for TGF which controls the activity of the Smad pathway. In the present study, researchers utilized a tetracycline inducible TIEG overexpressing breast cancer cell line and TIEG null mouse embryo fibroblasts (MEFs) to establish whether TIEG plays a central role in eliciting the anti-proliferative effects of TGF. It is similar to TGF treatment, TIEG overexpression significantly decreases cellular proliferation. Consistent with a role in regulating cellular proliferation, TIEG overexpression increases the expression of the cyclin dependent kinase inhibitor p21. Interestingly, while cellular proliferation of wild-type MEFs is inhibited by TGF, proliferation of TIEG null MEFs is stimulated by TGF. Furthermore, TIEG null MEF cells display a decrease in Smad dependent transcription with a concomitant prolonged increase in Smad7 expression compared to wild-type cells. These data strongly suggest that TIEG plays a central role in the anti-proliferative response to TGF and may explain how a loss of TIEG expression contributes to the development of cancer.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for TIEG1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TIEG1 present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for TIEG1 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 TIEG1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
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