Intra-Assay Precision | Inter-Assay Precision | |||||
---|---|---|---|---|---|---|
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 28 | 37 | 30 |
Mean | 78 | 180 | 695 | 79 | 185 | 730 |
Standard Deviation | 2.3 | 3.4 | 17.3 | 7.2 | 17.2 | 46.8 |
CV% | 2.9 | 1.9 | 2.5 | 9.1 | 9.3 | 6.4 |
Intra-Assay Precision | Inter-Assay Precision | |||||
---|---|---|---|---|---|---|
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 22 | 21 | 22 |
Mean | 88 | 198 | 816 | 97 | 209 | 826 |
Standard Deviation | 3 | 4.9 | 19.6 | 8.1 | 17.3 | 47 |
CV% | 3.4 | 2.5 | 2.4 | 8.4 | 8.3 | 5.7 |
The recovery of TGF-beta 1 spiked to levels throughout the range of the assay followed by activation was evaluated.
Sample Type | Average % Recovery | Range % |
---|---|---|
Cell Culture Supernates (n=4) | 99 | 86-115 |
Urine (n=3) | 96 | 88-100 |
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.
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