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EK000055-EK0448: Human MIP-1α ELISA Kit
Range 31.2pg/ml-2000pg/ml
Sensitivity < 10pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of human MIP-1α in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Macrophage inflammatory protein-1 alpha (MIP-1 alpha), also called CCL3, LD78. The cDNA for MIP-1 alpha predicts a mature peptide of 69 amino acids with a molecular mass of 7,889 daltons.1 LD78 is a member of a newly identified superfamily of small inducible proteins involved in inflammatory responses, wound healing, and tumorigenesis.2 MIP-1 alpha is a chemokine that has pro-inflammatory and stem cell inhibitory activities in vitro.3 It constitutes an important second signal for mast cell degranulation in the conjunctiva in vivo and consequently for acute-phase disease.4 The standard product used in this kit is recombinant human MIP-1a, consisting of 66 amino acids with the molecular mass of 7.5kDa.
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. Wolpe, S. D.; Cerami, A. Macrophage inflammatory proteins 1 and 2: members of a novel superfamily of cytokines. FASEB J. 3: 2565-2573, 1989.
2. Nakao, M.; Nomiyama, H.; Shimada, K. Structures of human genes coding for cytokine LD78 and their expression. Molec. Cell. Biol. 10: 3646-3658, 1990.
3. Cook, D. N.; Beck, M. A.; Coffman, T. M.; Kirby, S. L.; Sheridan, J. F.; Pragnell, I. B.; Smithies, O. Requirement of MIP-1-alpha for an inflammatory response to viral infection. Science 269: 1583-1585, 1995.
4. Miyazaki, D.; Nakamura, T.; Toda, M.; Cheung-Chau, K.-W.; Richardson, R. M.; Ono, S. J. Macrophage inflammatory protein-1-alpha as a costimulatory signal for mast cell-mediated immediate hypersensitivity reactions. J. Clin. Invest. 115: 434-442, 2005.
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