TGFbeta1 is secreted as a latent form, which consists of its mature form and a latency-associated peptide(beta1-LAP) in either the presence or the absence of additional latent TGF-beta1-binding protein. Processing and cleavage of the precursor protein between amino acids 278 and 279 results in the formation of LAP dimers and TGF beta dimers that then non-covalently associate with each other to form the small latent TGF beta complex. LAP is secreted and can be found in the extracellular matrix. In addition, LAP can also be expressed on platelets and activated regulatory T cells.
Sample Type:Cell culture supernates, serum and plasma (heparin, EDTA)
Intended Use:For quantitative detection of human LAP(TGF-beta1) in cell culture supernates, serum and plasma (heparin, EDTA).
Sensitivity:<10pg/ml
Range:62.5-4000pg/ml
Specificity:Natural and recombinant human LAP(TGF-beta1)
Test Principle:This human LAP(TGF-beta1) BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for LAP(TGF-beta1) has been precoated onto 96-well plates. Standards(sf21) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for LAP(TGF-beta1) is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human LAP(TGF-beta1) amount of sample captured in plate.
Kit Components:96-well plate precoated with human LAP(TGF-beta1) antibody, 1Lyophilized recombinant human LAP(TGF-beta1) standard, 10ng/tube×2Biotinylated human LAP(TGF-beta1) antibody, 130ul (dilution 1:100)Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100)Sample diluent buffer, 30mlAntibody diluent buffer, 12mlABC diluent buffer, 12mlTMB color developing agent, 10mlTMB stop solution, 10ml
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.
