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EK000029-EK0438: Mouse Leptin ELISA Kit
Range 62.5pg/ml-4000pg/ml
Sensitivity < 10pg/ml
Specificity: No detectable cross-reactivity with any other cytokine.
Application: For quantitative detection of mouse Leptin in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Leptin (or obese, OB) is a circulating hormone that is expressed abundantly and specifically in the adipose tissue.1 Human ob protein is a 166-amino acid polypeptide with a putative signal sequence and is 84 and 83% homologous to the mouse and rat ob proteins, respectively.2 Northern blot analysis reveals that OB RNA is present at a high level in adipose tissue but at much lower levels in placenta and heart.3 Indeed, Leptin is first described as an adipocyte-derived signalling factor, which, after interaction with its receptors, induces a complex response including control of bodyweight and energy expenditure. Leptin seems in addition to its role in metabolic control to have important roles in reproduction and neuroendocrine signalling.4 Leptin is a powerful inhibitor of bone formation in vivo.5 Comparative mapping of mouse and human DNA indicates that the ob gene is located within a region of mouse chromosome 6 that is homologous to a portion of human chromosome 7q.3 The standard product used in this kit is recombinant human Leptin.
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. Masuzaki, H.; Ogawa, Y.; Sagawa, N.; Hosoda, K.; Matsumoto, T.; Mise, H.; Nishimura, H.; Yoshimasa, Y.; Tanaka, I.; Mori, T.; Nakao, K. Nonadipose tissue production of Leptin: Leptin as a novel placenta-derived hormone in humans. Nature Med. 3: 1029-1033, 1997.
2. Masuzaki, H.; Ogawa, Y.; Isse, N.; Satoh, N.; Okazaki, T.; Shigemoto, M.; Mori, K.; Tamura, N.; Hosoda, K.; Yoshimasa, Y.; Jingami, H.; Kawada, T.; Nakao, K. Human obese gene expression: adipocyte-specific expression and regional differences in the adipose tissue. Diabetes 44: 855-858, 1995.
3. Green, E. D.; Maffei, M.; Braden, V. V.; Proenca, R.; DeSilva, U.; Zhang, Y.; Chua, S. C., Jr.; Leibel, R. L.; Weissenbach, J.; Friedman, J. M. The human obese (OB) gene: RNA expression pattern and mapping on the physical, cytogenetic, and genetic maps of chromosome 7. Genome Res. 5: 5-12, 1995.
4. Auwerx, J.; Staels, B. Leptin. Lancet 351: 737-742, 1998.
5. Elefteriou, F.; Takeda, S.; Ebihara, K.; Magre, J.; Patano, N.; Ae Kim, C.; Ogawa, Y.; Liu, X.; Ware, S. M.; Craigen, W. J.; Robert, J. J.; Vinson, C.; Nakao, K.; Capeau, J.; Karsenty, G. Serum Leptin level is a regulator of bone mass. Proc. Nat. Acad. Sci. 101: 3258-3263, 2004.
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