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EK000056-EK0453: Human MIP-3α ELISA Kit
Range 15.6pg/ml-1000pg/ml
Sensitivity < 1 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of human MIP-3α 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 3a (MIP3a), also called Chemokine, cc motif, ligand 20 (CCL20). The MIP-3alpha/CCL20 gene was cloned and sequenced, revealing a four exon, three intron structure, and was localized by FISH analysis to 2q35-q36.1 MIP3a is predominantly expressed in lymph nodes, appendix, PBL, fetal liver, fetal lung and several cell lines.2 MIP3a/CCL20 and its receptor CCR6 are markedly up-regulated in psoriasis, and they may play a role in the recruitment of T cells to lesional psoriatic skin.3 And Alanine MIP-3a and Serine MIP-3a, the two forms of MIP3a, that differ by one amino acid at the predicted signal peptide cleavage site. Both of them were chemically synthesized and tested for biological activity. And both flu antigen plus IL-2-activated CD4(+) and CD8(+) T lymphoblasts and cord blood-derived dendritic cells responded to Ser and Ala MIP-3a.1 The standard product used in this kit is recombinant human MIP-3a, consisting of 70 amino acids with the molecular mass of 8KDa.
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. Nelson, R. T.; Boyd, J.; Gladue, R. P.; Paradis, T.; Thomas, R.; Cunningham, A. C.; Lira, P.; Brissette, W. H.; Hayes, L.; Hames, L. M.; Neote, K. S.; McColl, S. R. : Genomic organization of the CC chemokine MIP-3-alpha/CCL20/LARC/EXODUS/SCYA20, showing gene structure, splice variants, and chromosome localization. Genomics 73: 28-37, 2001.
2. Rossi, D. L.; Vicari, A. P.; Franz-Bacon, K.; McClanahan, T. K.; Zlotnik, A. : Identification through bioinformatics of two new macrophage proinflammatory human chemokines. J. Immun. 158: 1033-1036, 1997.
3. Homey, B.; Dieu-Nosjean, M.-C.; Wiesenborn, A.; Massacrier, C.; Pin, J.-J.; Oldham, E.; Catron, D.; Buchanan, M. E.; Muller, A.; de Waal Malefyt, R.; Deng, G.; Orozco, R.; Ruzicka, T.; Lehmann, P.; Lebecque, S.; Caux, C.; Zlotnik, A. : Up-regulation of macrophage inflammatory protein-3-alpha/CCL20 and CC chemokine receptor 6 in psoriasis. J. Immun. 164: 6621-6632, 2000.
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