Rat 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. Standard, control, or sample and the working solution of Biotin-Conjugate are pipetted into the wells. Following incubation and wash steps, any TGF-β1 present is bound by the immobilized antibody and the detection antibody specific for TGF-β1 is 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 beta (TGF-β) proteins (including the three closely related mammalian isoforms TGF-β1, -2 and -3) are pleiotropic cytokines that regulate extracellular matrix production, wound healing, immune functions, cell proliferation and differentiation. Transforming growth factor beta (TGF-beta) is a TGF-beta superfamily that regulates cell growth and differentiation. Besides TGF-β1 to TGF-β5, this family also has activins, statins, Mullerian inhibitors (MIS) and bone morphogenetic proteins (BMPs). TGF-β is named after the cytokine converts the phenotype of normal fibroblasts.

Tissues that are generally active in cell differentiation often contain higher levels of TGF-β, such as hematopoietic cells of osteoblasts, kidneys, bone marrow, and fetal liver. The level of TGF-β1 is the highest in human platelets and mammalian bone. TGF-β1 is present in an inactive state and activated when TGF-β1 is partially released. TGF-β1 has dual functions of activation and inhibition. Many TGF-β1 receptors regulate their biological activities, including receptor I (53-65KD), receptor II (83-110KD), receptor III (250-310KD), receptor IV (60KD), and receptor V (400KD).

TGF-β is a dimer formed by two disulfide bonds of the same or similar molecular weight of 12.5 kDa. The rat TGF-β1 precursor cDNA encodes a 390 amino acid residue comprising a 29 amino acid residue signal peptide and a 361 amino acid residue precursor protein. The precursor protein is hydrolyzed by furin to form a potentially related peptide with an N-terminal 249 amino acid residue and a C-terminal 112 amino acid residue mature TGF-β1.

The biological function of TGF-β is mainly in inflammation, tissue repair and embryo development. TGF-β plays an important role in regulating cell growth, differentiation and immune function. TGF-β can inhibit the proliferation of immunocompetent cells, lymphocyte differentiation and cytokine production (such as IFN-γ and TNF-α production in PBMC).

GENE ID 59086
SWISS PROT Q63264
SYNONYMS Tgfb

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with antibody to rat TGF-β1 (812) 1 plate
Rat TGF-β1 standard lyophilized, 4000pg/ml upon reconstitution 2 vials
Concentrated Biotin-Conjugate anti-rat TGF-β1 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 62.50 - 4000 pg/mL
SPECIFICITY This assay recognizes both natural and recombinant Rat TGF-β1. The factors listed below were prepared at 50ng/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, BMP-2, BMP-3, BMP-3b, BMP-4, BMP-5, BMP-6, BMP-8b, BMP-10, BMP-15, LAP, BMPR-IA, BMPR-IB, BMPR-II, Follistatin288, Follistatin300, Follistatin315, TGF-βRI, TGF-βRII, TGF-βRIII
Recombinant mouse BMPR-IA, BMPR-IB, BMP-3b, TGF-βRI