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EK000097-EK0500: Mouse SDF-1 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 SDF-1 in plasma, body fluids, tissue lysates or cell culture supernates.
Background
SDF-1 (stromal cell-derived factor-1) is small cytokine belonging to the chemokine family that is officially designated Chemokine (C-X-C motif) ligand 12 (CXCL12). This gene is located on chromosome 10q11.1.1 SDF-1 is produced in two forms, SDF-1a/CXCL12a and SDF-1?/CXCL12b, by alternate splicing of the same gene.2 Chemokines are characterized by the presence of four conserved cysteines, which form two disulfide bonds. The CXCL12 proteins belong to the group of CXC chemokines, whose initial pair of cysteines are separated by one intervening amino acid. CXCL12 is strongly chemotactic for lymphocytes.3,4,5,6 CXCL12 was shown to be expressed in many tissues in mice (including brain, thymus, heart, lung, liver, kidney, spleen and bone marrow). CXCL12 is a highly efficacious lymphocyte chemoattractant. In addition, CXCL12 induces intracellular actin polymerization in lymphocytes.7 CXCL12 is a substrate for the matrix metalloproteinase-2, which cleaves an CXCL12 N-terminal tetrapeptide.8 The standard product used in this kit is recombinant SDF-1 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. Shirozu, M.; Nakano, T.; Inazawa, J.; Tashiro, K.; Tada, H.; Shinohara, T.; Honjo, T. : Structure and chromosomal localization of the human stromal cell-derived factor 1 (SDF1) gene. Genomics 28: 495-500, 1995.
2. De La Luz Sierra et al. Differential processing of stromal-derived factor-1alpha and beta explains functional diversity. Blood 103:2452-2459, 2004.
3. Bleul et al. A highly efficacious lymphocyte chemoattractant, stromal cell-derived factor 1 (SDF-1). J. Exp. Med. 184: 1101-1109, 1996.
4. Ara et al. Impaired colonization of the gonads by primordial germ cells in mice lacking a chemokine, stromal cell-derived factor-1 (SDF-1). Proc. Nat. Acad. Sci. 100: 5319-5323, 2003.
5. Askari et al. Effect of stromal-cell-derived factor 1 on stem-cell homing and tissue regeneration in ischaemic cardiomyopathy. Lancet 362: 697-703, 2003.
6. Ma et al. Impaired B-lymphopoiesis, myelopoiesis, and derailed cerebellar neuron migration in CXCR4- and SDF-1-deficient mice. Proc. Nat. Acad. Sci. 95: 9448-9453, 1998.
7. Bleul, C. C.; Fuhlbrigge, R. C.; Casasnovas, J. M.; Aiuti, A.; Springer, T. A. : A highly efficacious lymphocyte chemoattractant, stromal cell-derived factor 1 (SDF-1). J. Exp. Med. 184: 1101-1109, 1996.
8. McQuibban, G. A.; Butler, G. S.; Gong, J. H.; Bendall, L.; Power, C.; Clark-Lewis, I.; Overall, C. M. : Matrix metalloproteinase activity inactivates the CXC chemokine stromal cell-derived factor-1. J. Biol. Chem. 276: 43503-43508, 2001.
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