Description

Nori Feline TGFb1 ELISA Kit Summary

Alternative names for TGFb1: Transforming growth factor beta 1

Alternative names for feline: cat

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background:

TGFβ1 (transforming growth factor beta 1) was first identified in human platelets as a protein with a molecular mass of 25 kilodaltons with a potential role in wound healing (1).  TGFB1, TGFB2, and TGFB3 all function through the same receptor signaling systems. They are members of the large TGFβ superfamily. TGFβ proteins are highly pleiotropic cytokines that regulate processes such as immune function, proliferation and epithelial mesenchymal transition (2-4). It was later characterized as a large protein precursor (containing 390 amino acids) that was proteolytically processed to produce a mature peptide of 112 amino acids (5). TGFβ activation from latency is controlled both spatially and temporally, by multiple pathways that include actions of proteases such as plasmin and MMP9, and/or by thrombospondin 1 or selected integrins (5, 6). Although different isoforms of TGFβ are naturally associated with their own distinct LAPs, the TGFβ1 LAP is capable of complexing with, and inactivating, all other Equine TGFβ isoforms and those of most other species (7). Mutations within the LAP are associated with Camurati Engelmann disease, a rare sclerosing bone dysplasia characterized by inappropriate presence of active TGFβ1 (8).

References

1. Assoian R, et al. (1983). J Biol Chem 258 (11): 7155.
2. Dunker, N. and K. Krieglstein (2000) Eur. J. Biochem.267:6982.
3. Wahl, S.M. (2006) Immunol. Rev. 213:213.
4. Chang, H. et al. (2002) Endocr. Rev. 23:787.
5. Derynck, R. et al. (1985) Nature 316:701.
6. Oklu, R. and R. Hesketh (2000) Biochem. J. 352:601.
7. Miller, D.M. et al. (1992) Mol. Endocrinol. 6:694.
8. Janssens, K. et al. (2003) J. Biol. Chem. 278:7718.

Product Citation