Mouse TGF-β1 ELISA
The mTGF-β1 ELISA is an enzyme-linked immunosorbent assay for the quantitative detection of mouse transforming growth factor beta-1 levels in cell culture supernatants, serum, plasma, or other body fluids.
TGF-β1 is the first recognized transforming growth factor, its subunits of each 12.5 kDa are bound via disulphide bridges. TGF-β1 is inhibitive to T- and B cell proliferation as well as to maturation and activation of macrophages. It furthermore inhibits activity of natural killer cells and lymphokine activated killer cells and blocks production of cytokines.
An anti-mTGF-β1 coating antibody is adsorbed onto microwells.
mTGFβ1 present in the sample or standard binds to antibodies adsorbed to the microwells; a biotin-conjugated monoclonal anti-mTGFβ1 antibody is added and binds to mTGFβ1 captured by the first antibody.
Following incubation unbound biotin conjugated antimTGFβ1 is removed during a wash step. Streptavidin-HRP is added and binds to the biotin conjugated anti-mTGFβ1 Following incubation unbound Streptavidin-HRP is removed during a wash step, and substrate solution reactive with HRP is added to the wells.
A coloured product is formed in proportion to the amount of mTGFβ1 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven mTGFβ1 standard dilutions and mTGFβ1 sample concentration determined.