TGF-Beta 1 ELISA Assay

$445.00

The Eagle Biosciences Human TGF Beta 1 (TGF-β1) ELISA Assay Kit (enzyme-linked immunoassay kit) is intended for the quantitative determination of human TGF Beta 1 (TGF-β1) concentrations in cell culture supernates, serum, and plasma. The Eagle Biosciences Human TGF Beta 1 (TGF-β1) ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: TB131-K01 Categories: , ,

TGF-Beta 1 ELISA Assay

The TGF-Beta 1 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 15 pg/mL
Dynamic Range: 62.5 – 1000 pg/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Names: Transforming Growth Factor


Assay Background

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.


Related Products

TNF-Alpha ELISA Assay Kit
Human G-CSF ELISA Assay
Human EGF ELISA Assay Kit

Additional Information

Assay Principle


The Eagle Biosciences Human TGF Beta 1 (TGF-β1) ELISA Assay Kit 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 colored 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.

  1. Prepare all reagents and working standards as directed in the previous sections.
  2. Determine the number of microwell strips required to test the desired number of samples plus appropriate number of wells needed for running blanks and standards. Remove extra microwell strips from holder and store in foil bag with the desiccant provided at 2-8°C sealed tightly.
  3. Add 100 µlof Standard, control, or sample, per well. Cover with the adhesive strip provided. Incubate for 1.5 hours at 37C
  4. Aspirate each well and wash, repeating the process three times for a total of four washes. Wash by filling each well with Wash Buffer (350 µl) using a squirt bottle, manifold dispenser or auto-washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
  5. Add 100 µl of the working solution of Biotin-Conjugate to each well. Cover with a new adhesive strip and incubate 1 hour at 37C
  6. Repeat the aspiration/wash.
  7. Add 100 µL of the working solution of Streptavidin-HRP to each well. Cover with a new adhesive strip and incubate for 30 minutes at 37C.   Avoid placing the plate in direct light.
  8. Repeat the aspiration/wash.
  9. Add 100 µL of Substrate Solution to each well. Incubate for 10-20 minutes at 37C. Avoid placing the plate in direct light.
  10. Add 100 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  11. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. (optionally 630nm as the reference wave length;610-650nm is acceptable)

Documents

Product Manual


 

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

Publications

References


1.    J Biol Chem. 2001 Dec 17
2.    Cell Signal. 2002 Jan;14(1):31-36.
3.    Glia. 2001 Dec;36(3):354-363.
4.    J Cell Biochem. 2001;81(S36):79-88.
5.    J Neurosci Res. 2001 Nov 1;66(3):369-376

Product Citations