预期应用
ELISA法定量测定猪血清、血浆、细胞培养物上清或其它相关液体中TGFβ1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TGFβ1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TGFβ1抗原、生物素化的抗猪TGFβ1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TGFβ1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 porcine TGFβ1 concentrations in cell culture supernates, serum, and plasma.
Introduction
TGF-β is capable of producing a variety of effects and virtually all cell types respond to this factor in some way. The inappropriate presence of active TGF-β1 has been implicated in a variety of pathological conditions Because of the necessity for regulating its activity tightly, TGF-β is secreted by cells in the form of an inactive complex. This complex consists of TGF-β1 associated non-covalently with a protein designated the latency associated peptide (LAP). TGF-β1 and LAP represent components of a pro-peptide that is cleaved in a post-golgi compartment prior to secretion. LAP and TGF-β1 each consist of a disulfide-linked homodimer and the association of these two components renders TGF-β1 inactive and inaccessible to anti-TGF-β antibodies. In many instances in vivo, associated LAP and TGF-β, called the small latent TGF-β complex, are bound to an additional protein designated the latent TGF-β1 binding protein (LTBP), forming the large latent TGF-β1 complex. The mechanisms by which active TGF-β1 is released from these latent complexes represent important control steps for the regulation of the numerous effects of TGF-β1. Despite considerable study, these mechanisms are not well understood.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for TGFβ1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TGFβ1 present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for TGFβ1 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 TGFβ1 bound in the initial step. The color
development is stopped and the intensity of the color is measured.
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