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EK000052-EK0520: Human TIMP-1 ELISA Kit
Range 156pg/ml-10,000pg/ml
Sensitivity < 5 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitative detection of human TIMP-1 in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
The tissue inhibitor of metalloproteinases 1 (TIMP1) is also called erythroid-potentiating activity (EPA). The X-linked gene for human TIMP1 is expressed in some but not all inactive X-containing somatic-cell hybrids, suggesting that this gene is either prone to reactivation or variable in its inactivation.1 Purified EPA specifically stimulates human and murine cells of the erythroid lineage, unlike murine interleukin-3 (IL-3) which stimulates precursor cells from all haematopoietic lineages.2 TIMP1 is thought to play a regulatory role in connective tissues by forming inactive complexes with those metalloproteinases that are normally responsible for connective tissue turnover. The human gene encoding TIMP has been mapped to the X chromosome in the region Xp11.1-p11.4.3 The standard product used in this kit is natural TIMP-1 with the molecular mass of 22KDa.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Anderson, C. L.; Brown, C. J. Polymorphic X-chromosome inactivation of the human TIMP1 gene. Am. J. Hum. Genet. 65: 699-708, 1999.
2. Gasson, J. C.; Golde, D. W.; Kaufman, S. E.; Westbrook, C. A.; Hewick, R. M.; Kaufman, R. J.; Wong, G. G.; Temple, P. A.; Leary, A. C.; Brown, E. L.; Orr, E. C.; Clark, S. C. Molecular characterization and expression of the gene encoding human erythroid-potentiating activity. Nature 315: 768-771, 1985.
3. Spurr, N. K.; Goodfellow, P. N.; Docherty, A. J. P. Chromosomal assignment of the gene encoding the human tissue inhibitor of metalloproteinases to Xp11.1-p11.4. Ann. Hum. Genet. 51: 189-194, 1987.
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