Human TGF-β1 ELISA Kit

Principle of the Assay

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for TGF-β1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TGF-β1 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for TGF-β1 is added to the wells and binds to the combination of capture antibody- TGF-β1 in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of TGF-β1 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-β1 standard dilutions and TGF-β1 sample concentration determined.

For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.

Target Information

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-α and TGF-β. These molecules are members of a superfamily that includes TGF-β1 through 5, bone morphogenic proteins, activins and inhibins. It plays a critical role in cellular growth, development, differentiation, proliferation, extracellular matrix (ECM) synthesis and degradation, control of mesenchymal-epithelial interactions during embryogenesis, immune modulation, apoptosis, cell cycle progression, angiogenesis, adhesion and migration and leukocyte chemotaxis.

Originally, TGF-β1 was separated from platelets and later found that TGF-β1 can be expressed in many organizations. Human TGF-β1 is a 25kDa, disulfide-linked, non-glycosylated homodimer. Biological activity of TGF-β1 is regulated by a number of receptors, including receptor I (53-65KD), receptor II (83-110KD), receptor III (250-310KD), receptor-IV (60KD) and receptor V (400KD).

TGF-β1 is the key mediator in the pathophysiology of tissue repair and human fibrogenesis: balance between production and degradation of type I collagen, and fibrosis and scarring in organ and tissue. TGF-β1 exhibits important immunoregulatory features of partially adverse character: TGF-β1 inhibits B and T cell proliferation, differentiation and antibody production as well as maturation and activation of macrophages. TGF-β1 is synthesized, with only a few exceptions, by virtually all cells, and TGF receptors are expressed by all cells. TGF-β affects nearly every physiological process in some way; its systemic and cell-specific activities are too complicated to review here. There are, however, three fundamental activities: TGF-β1 modulates cell proliferation, generally as a suppressor; TGF-β1 enhances the deposition of extracellular matrix through promotion of synthesis and inhibition of degradation; TGF-β1 is immunosuppressive through a variety of mechanisms. The specific action of TGF-β on a particular cell depends on the exact circumstances of that cell’s environment.

GENE ID 7040
SWISS PROT P01137
SYNONYMS CED; LAP; DPD1; TGFB; IBDIMDE; TGFbeta

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human TGF-β1 (8﹡12) 1 plate
Human TGF-β1 Standard lyophilized, 4000 pg/ml upon reconstitution 2 vials
Concentrated Biotin-Conjugate anti-human TGF-β1 monoclonal antibody 2 vials
Streptavidin-HRP solution 2 vials
Standard /sample Diluent 4 bottles
Biotin-Conjugate antibody Diluent 1 bottle
Streptavidin-HRP Diluent 1 bottle
Wash Buffer Concentrate 20x (PBS with 1% Tween-20) 1 bottle
Substrate Solution 1 vial
Stop Solution 1 vial
Adhesive Films 4 pieces
Product data sheet 1 copy

Storage

Storage Store at 2 - 8°C

Performance Characteristics

REPEATABILITY The coefficient of variation of both intra-assay and inter-assay were less than 10%.
SENSITIVITY The minimum detectable dose was 31pg/mL.
ASSAY RANGE 62.5 - 4000 pg/mL
SPECIFICITY This assay recognizes both natural and recombinant human TGF-β1. The factors listed below were prepared at 100ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.
Factors assayed for cross-reactivity
Recombinant human Activin A, Activin RIA, Activin RIIA, Activin RIIB, BMP-2, BMP-3, BMP-3b, BMP-4, BMP-5, BMP-6, BMP-8b, BMP-10, BMP-15, BMP-2, TGF-β2, TGF-β3, TGF-α
Recombinant mouse BMP-3b, BMPR-IA , BMPR-IB, TGF-β RI
Other proteins Natural porcine TGF-β2