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EK000045-EK0517: Mouse TPO ELISA Kit
Range 31.2pg/ml-2000pg/ml
Sensitivity < 5 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitative detection of mouse TPO in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Thrombopoietin (TPO) is also called megakaryocyte growth and development factor (MGDF). TPO, a lineage-specific cytokine affecting the proliferation and maturation of megakaryocytes from committed progenitor cells, is believed to be the major physiological regulator of circulating platelet levels.1 Human, dog, and mouse cDNAs for MGDF are highly conserved and encode open reading frames for proteins of 353, 352, and 356 amino acids, respectively, including predicted signal peptides. MGDF is a novel cytokine that regulates megakaryocyte development and is a ligand for the MPL receptor. The human MGDF gene has been mapped to chromosome 3q26.3. 2,3 The standard product used in this kit is gene recombinant mouse TPO, consisting of 335 amino acids with the molecular mass of 35KDa. As a result of glycosylation, the molecular mass is 75KDa.
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. Foster, D. C.; Sprecher, C. A.; Grant, F. J.; Kramer, J. M.; Kuijper, J. L.; Holly, R. D.; Whitmore, T. E.; Heipel, M. D.; Bell, L. A.; Ching, A. F. T.; McGrane, V.; Hart, C.; O'Hara, P. J.; Lok, S. Human thrombopoietin: gene structure, cDNA sequence, expression, and chromosomal localization. Proc. Nat. Acad. Sci. 91: 13023-13027, 1994.
2. Bartley, T. D.; Bogenberger, J.; Hunt, P.; Li, Y.-S.; Lu, H. S.; Martin, F.; Chang, M.-S.; Samal, B.; Nichol, J. L.; Swift, S.; Johnson, M. J.; Hsu, R.-Y.; and 41 others. Identification and cloning of a megakaryocyte growth and development factor that is a ligand for the cytokine receptor Mpl. Cell 77: 1117-1124, 1994.
3. Chang, M.; McNinch, J.; Basu, R.; Shutter, J.; Hsu, R.; Perkins, C.; Mar, V.; Suggs, S.; Welcher, A.; Li, L.; Lu, H.; Bartley, T.; Hunt, P.; Martin, F.; Samal, B.; Bogenberger, J. Cloning and characterization of the human megakaryocyte growth and development factor (MGDF) gene. J. Biol. Chem. 270: 511-514, 1995.
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