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EK000016-EK0403: Mouse IL-3 ELISA Kit
Range 7.8pg/ml-500pg/ml
Sensitivity < 1 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of mouse IL-3 in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
The gene IL-3 encodes interleukin 3, a hematopoietic colony-stimulating factor (CSF) that is capable of supporting the proliferation of a broad range of hematopoietic cell types.1 Interleukin-3 (IL-3), a protein of 140 amino acids, is chemically synthesized by means of an automated peptide synthesizer and is shown to have the biological activities attributed to native IL-3.2 The cDNA sequence for murine interleukin-3, one of the colony stimulating factors that regulate haematopoiesis, codes for a polypeptide of 166 amino acids including a putative signal peptide.3 The mouse IL 3 gene is located on chromosome 11.4 The human gene encoding IL 3 is tandemly arrayed on the long arm of chromosome 5.5 The standard product used in this kit is recombinant human IL-3, consisting of 133 amino acids with the molecular mass of 15KDa.
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. Le Beau, M. M.; Epstein, N. D.; O'Brien, S. J.; Nienhuis, A. W.; Yang, Y.-C.; Clark, S. C.; Rowley, J. D. The interleukin 3 gene is located on human chromosome 5 and is deleted in myeloid leukemias with a deletion of 5q. Proc. Nat. Acad. Sci. 84: 5913-5917, 1987.
2. Clark-Lewis, I.; Aebersold, R.; Ziltener, H.; Schrader, J. W.; Hood, L. E.; Kent, S. B. H. Automated chemical synthesis of a protein growth factor for hemopoietic cells, interleukin-3. Science 231: 134-139, 1986.
3. Fung, M. C.; Hapel, A. J.; Ymer, S.; Cohen, D. R.; Johnson, R. M.; Campbell, H. D.; Young, I. G. Molecular cloning of cDNA for murine interleukin-3. Nature 307: 233-237, 1984.
4. Ihle, J. N.; Silver, J.; Kozak, C. A. Genetic mapping of the mouse interleukin 3 gene to chromosome 11. J. Immun. 138: 3051-3054, 1987.
5. Yang, Y.-C.; Kovacic, S.; Kriz, R.; Wolf, S.; Clark, S. C.; Wellems, T. E.; Nienhuis, A.; Epstein, N. The human genes for GM-CSF and IL3 are closely linked in tandem on chromosome 5. Blood 71: 958-961, 1988.
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