Human/Mouse/Rat/Porcine/Canine TGF-β1 enzyme-linked immunoassay kit

CAT: EH0012 Datasheet
Specification 96 Test
Sensitivity 33.32 pg/ml (50 μl)
Standard Curve Range 156.25~10000 pg/ml
Standard Curve Gradient 7 Points/2 Folds
Number of Incubations 2
Sample Volume 50 μl
Type Ready-to-Use
Operation Duration 120min
pg/ml O.D. Average Corrected
0.00 0.0886 0.0786 0.0836
156.25 0.1488 0.1692 0.1590 0.0750
312.50 0.2321 0.2237 0.2279 0.1439
625.00 0.3551 0.3739 0.3645 0.2805
1250.00 0.5891 0.6252 0.6072 0.5232
2500.00 1.1123 1.1675 1.1399 1.0559
5000.00 1.9813 2.0695 2.0254 1.9414
10000.00 3.3531 3.2747 3.3139 3.2299

Precision

Intra-assay Precision Inter-assay Precision
Sample Number S1 S2 S3 S1 S2 S3
22 22 22 6 6 6
Average(pg/ml) 195.0 1090.8 3619.9 237.3 1154.4 3711.7
Standard Deviation 10.3 67.7 202.6 16.3 67.5 144.7
Coefficient of Variation(%) 5.3 6.2 5.6 6.8 5.8 3.9

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays) Three samples of known concentration were tested six times on one plate to assess intra-assay precision.

Spike Recovery

The spike recovery was evaluated by spiking 3 levels of mammal TGF beta1 into health serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 88% to 105% with an overall mean recovery of 99%.

Sample Values

Sample Matrix Sample Evaluated Range (ng/ml) Detectable (%) Mean of Detectable (ng/ml)
Serum 30 35.3-74.7 100 58.4

Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of TGF-β1 in this assay. No medical histories were available for the donors.

Background: TGF-β1

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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