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.
Transforming Growth Factor-beta (TGF-β) is a pleiotropic cytokine which exists in five isoforms, known as TGF-β1-5. TGF-β1 is the most abundant form in lymphoid organs and is found almost ubiquitously while other isoforms are expressed in a more restricted distribution. The biologically active forms of all isoforms are disulfide-linked homodimers. The heat- and acid- stable monomeric subunits have a length of 112 amino acids. TGF-β1 is produced in very high levels by platelets. Other cellular sources of TGF-β1 include macrophages, lymphocytes, endothelial cells, chondrocytes, and leukemic cells. TGF-β1 secretion can be induced by steroids, retinoids, EGF, NGF, vitamin D3, and IL-1. Activities of TGF-β1 include inhibition of cell growth for inhibitor for normal and transformed epithelial cells, endothelial cells, fibroblasts, neurons, and lymphoid cells and other hematopoietic cell types. TGF-β1 inhibits the proliferation of T cells and NK cells and down-regulates the activities of activated macrophages. TGF-β1 blocks the anti-tumor activity of IL-2–bearing lymphokine-activated killer (LAK) cells. TGF-β1 has a critical role in the development of regulatory T cells. Dendritic cells exposed to tumors have been reported to secrete TGF-β1 and stimulate expansion of naturally-occurring T reg cells. Moreover, TGF-β1 has been shown to act as a costimulatory factor for expression of Foxp3, leading to the differentiation of CD4+CD25+ Treg cells from peripheral CD4+CD25- progeny. TGF-β-induced regulatory T cells have been termed Ti-Treg.
|SYNONYMS||TGF-beta1, TGFbeta1, Tgfb, Tgfb-1|
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to mouse TGF-β1 (8﹡12)||1 plate|
|Mouse TGF-β1 Standard lyophilized, 1000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-mouse 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|
|1N HCl||1 vial|
|1 N NaOH||1 vial|
|2.7N NaOH||2 vials|
|2.5N Acetic Acid/10 M Urea||2 vials|
|Adhesive Films||4 pieces|
|Product data sheet||1 copy|
|Storage||Store at 2 - 8°C|
|REPEATABILITY||The coefficient of variation of both intra-assay and inter-assay were less than 10%.|
|SENSITIVITY||The minimum detectable dose was 7pg/mL.|
|ASSAY RANGE||15.6-1000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant mouse 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||TGF-β2, TGF-β3|
|Recombinant mouse||TGF-β RI/Fc Chimera|
Data Analysis Assistance
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