Transforming growth factor beta 1 or TGFβ-1 is a human polypeptide encoded by the TGFB1 gene. This gene is on the q arm of chromosome 19. TGFβ-1 is a cytokine and belongs to the transforming growth factor beta superfamily. TGFβ-1 regulates several cellular functions, including cell proliferation, cell growth, cell differentiation, cell motility, and apoptosis. TGFβ-1 is present throughout the body but abundantly found in skeletal tissues regulating bone and cartilage formation. Researchers had first discovered TGFβ-1 as a 25 kilodaltons protein in the human platelets. It showed potential in healing wounds. Later, the researchers distinguished it into a 390 amino acid precursor protein that converts by proteolytic cleavage into a 112 amino acid mature peptide.
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We use Luminex, ELISA, and MSD assays to detect TGFβ-1 in serum, plasma, and tissue/cell culture samples. Besides the criteria listed in the protocol, we also test dilution linearity, cross-reactivity, kit stability, and sample detectability.
Following is an elaborated Luminex protocol.
We begin with 200 μl of assay buffer in each well and shake it for 10 mins at RT. We decant the assay buffer and residues and introduce 25 μl of the following ingredients in appropriate wells: Standards, control or treated samples, followed by assay buffer, matrix solution, , and beads and incubate it at RT for 2hrs or overnight at 4 °C. After incubation, plates are incubated for 2 mins on magnetic plate, following which well contents are removed and washed with a 200 μl wash buffer. Once the beads capture a sample analyte, we incubate them with a biotinylated detection antibody. We then complete the reaction by incubating with 25 μl Streptavidin-Phycoerythrin, a reporter molecule in each well for 30 mins at RT. Post-incubation, we remove the well contents and wash them with a 200 μl wash buffer and add 100 μl of drive or sheath fluid. We read the assay on an appropriate Luminex platform.
The transforming growth factor beta 1 test or assay identifies each microsphere through fluorescent reporter signals. We can perform this assay in a single day or over two days with overnight incubation and it requires 25 μl of 1:30 dilution serum/plasma post-treatment. The assay has sensitivity between 8-12 pg/ml and an accuracy of 62-128 %.
TGFβ-1 plays a crucial role in human development. The pathways associated with TGFβ-1 signaling are complex and finely orchestrated through intertwined channels with several signaling pathways. In mouse models, studies have located TGFβ-1 mRNA and/or protein in the cartilage, endochondral, and skin, suggesting its role in the differentiation and growth of these tissues.
Learn more about TGFβ-1 and its functions in the following resources.
https://ghr.nlm.nih.gov/gene/TGFB1
https://www.ncbi.nlm.nih.gov/gene/7040
https://translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-10-183
https://www.uniprot.org/uniprot/P01137
https://www.genecards.org/cgi-bin/carddisp.pl?gene=TGFB1
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TGFβ-1 is a multifunctional protein. It synergistically interacts with TGFA and induces transformation, and antagonistically acts as an autocrine growth factor. TGF-β abnormality may induce apoptosis. Through specialized receptors, TGFβ-1 interacts uniquely with different cell types and developmental stages. Most leukocytes produce TGFβ-1. Regulatory T cells secrete TGFβ-1 to inhibit IL- 1/2 dependent T cell proliferation and activate cytotoxic and helper T cells. TGFβ-1 can decrease IL-2 receptor expression to down-regulate immune cell activity and increase specific T cells expression to proliferate immature cells. Similarly, TGFβ-1 inhibits B cell proliferation and stimulates apoptosis, and regulates protein expression of mature and immature B cells.
TGFβ-1 has a suppressive and sometimes opposite effect on myeloid cells. TGFβ-1 may inhibit macrophage and monocyte proliferation and intercept the production of reactive nitrogen and oxygen intermediates. TGFβ-1 works as a chemoattractant. Low levels of TGFβ-1 attract monocytes and macrophages. Thus, TGFβ-1 lowers its levels to elicit an immune response chemotactically.