Transforming growth factor (TGF), a ‘factor’ that promoted the transformation of cultured fibroblasts into a tumor-like phenotype, was subsequently found to be more of a tumor suppressor than tumor promoter and to be a mixture of two proteins, TGF-alpha and TGF-beta. Human TGF-beta1 is a 25kD, disulfide-linked, non-glycosylated homodimer. There is nearly 100% sequence conservation across mammalian species. Biological activity requires release of TGF-beta1 from the latent complex. This can be done in vitro by disruption of LAP (e.g. acidification). There are, however, three fundamental activities: TGF-beta1 modulates cell proliferation, generally as a supressor; TGF-beta1 enhances the deposition of extracellular matrix through promotion of synthesis and inhibition of degradation; TGF-beta1 is immunosuppressive through a variety of mechanisms. The specific action of TGF-beta on a particular cell depends on the exact circumstances of that cell’s environment.
Sample Type:Cell culture supernatants, serum, plasma, and tissue
Intended Use:The Human TGF-beta1 BioAssay™ ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human TGF-beta1 concentrations in cell culture supernatants, serum, plasma, and tissue.
Sensitivity:7pg/ml
Range:31.25-2000pg/ml
Specificity:Recognizes human TGF-beta1. There was no detectable cross reactivity with any of the tested proteins following IL-1,2,3,4,5,6, 8,10,12,TNF,VEGF,ICAM and IFN.
Sample Volume:100ul/well
Test Principle:This assay employs the Sandwich immunoassay technique. An anti-h TGF-beta1 monoclonal coating antibody is adsorbed onto microwells. TGF-beta1 present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-h TGF-beta1 antibody is added and binds to TGF-beta1 captured by the first antibody. Following incubation unbound Biotinylated anti-h TGF-beta1 antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h TGF-beta1 antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of TGF-beta1 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 TGF-beta1 standard dilutions and TGF-beta1 sample concentration determined.
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
