• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Sample Type & Volume Required

  100 µL

• Range

  31.20 - 2,000 pg/mL

• Sufficient Materials

  For fifteen 96-well plates*

• Specificity

  Please see the

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

Ancillary Reagent Kit Available  

DY010, Sample Activation Kit 1 - DY007, DuoSet ELISA Ancillary Reagent Kit 1 -

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant TGF-ß1. The Reagent Diluent recommended may be suitable for most cell culture supernate, serum, and plasma samples. The Reagent Diluent selected for use can alter the performance of an immunoassay. Reagent Diluent optimization for samples with complex matrices such as serum and plasma, may improve their performance in this assay.

Product Features

• Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time

• Development protocols are provided to guide further assay optimization

• Assay can be customized to your specific needs

• Economical alternative to complete kits

Kit Content

• Capture Antibody

• Detection Antibody

• Recombinant Standard

• Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 1 (5 plates): (Catalog # ) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 1.

The components listed above may be purchased separately:
PBS: (Catalog # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # ), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # ), or 1.4% delipidized bovine serum, 0.05% Tween 20 in PBS, pH 7.2-7.4, 0.2 μm filtered.

Blocking Buffer: (Catalog # ), or 5% Tween 20 in PBS, pH 7.2-7.4, 0.2 μm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H₂O₂) and Color Reagent B (Tetramethylbenzidine)

Stop Solution: 2 N H₂SO₄ (Catalog # )

Microplates: R&D Systems (Catalog # )

Plate Sealers: ELISA Plate Sealers (Catalog # )

Activation Reagents Required:

Sample Activation Kit 1: 3 vials (10 mL/vial) of 1N HCL and 3 vials (10 mL/vial) of 1.2 N NaOH/1M HEPES (R&D Systems, Catalog # ).

Preparation and Storage

• Storage

  Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: TGF-beta 1

Transforming Growth Factor Beta 1, 2, and 3 (TGF-beta 1, TGF-beta 2, and TGF-beta 3) are highly pleiotropic cytokines that virtually all cell types secrete. TGF-beta molecules are proposed to act as cellular switches that regulate processes such as immune function, proliferation, and epithelial-mesenchymal transition. Targeted deletions of these genes in mice show that each TGF-beta isoform has some non-redundant functions: TGF-beta 1 is involved in hematopoiesis and endothelial differentiation; TGF-beta 2 affects development of cardiac, lung, craniofacial, limb, eye, ear, and urogenital systems; and TGF-beta 3 influences palatogenesis and pulmonary development. The full range of in vitro biological activities of TGF-beta 5 has not yet been explored. However, TGF-beta 1, TGF-beta 2, TGF-beta 3, and TGF-beta 5 have been found to be largely interchangeable in an inhibitory bioassay, and it is anticipated that TGF-beta 5 will show a spectrum of activities similar to the other TGF-beta family members. To date, the production of TGF-beta 5 has only been demonstrated in Xenopus.

TGF-beta ligands are initially synthesized as precursor proteins that undergo proteolytic cleavage. The mature segments form active ligand dimers via a disulfide-rich core consisting of the characteristic 'cysteine knot'. TGF-beta signaling begins with binding to a complex of the accessory receptor betaglycan (also known as TGF-beta RIII) and a type II serine/threonine kinase receptor termed TGF-beta RII. This receptor then phosphorylates and activates a type I serine/threonine kinase receptor, either ALK-1 or TGF-beta RI (also called ALK-5). The activated type I receptor phosphorylates and activates Smad proteins that regulate transcription. Use of other signaling pathways that are Smad-independent allows for distinct actions observed in response to TGF-beta in different contexts.

• Entrez Gene IDs:

  7040 (Human); 21803 (Mouse); 59086 (Rat); 397078 (Porcine); 100033900 (Equine);

• Long Name:

  Transforming Growth Factor beta 1

• Aliases:

  CEDLAP; DPD1; latency-associated peptide; TGFB; TGFB1; TGF-beta 1 protein; TGFbeta 1; TGF-beta 1; TGFbeta; TGF-beta-1; transforming growth factor beta-1; transforming growth factor, beta 1