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. Interleukin-3(IL-3), a protein of 140aa, is chemically synthesized by means of an automated peptide synthesizer and is shown to have the biological activities attributed to native IL-3. The cDNA sequence for murine interleukin-3, one of the colony stimulating factors that regulate haematopoiesis, codes for a polypeptide of 166aa including a putative signal peptide. The mouse IL 3 gene is located on chromosome 11. 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 133aa with the molecular mass of 15KDa.
Sample Type
Cell culture supernates and serum.
Intended Use
For quantitative detection of rat IL-3 in cell culture supernates and serum.
Specificity
Natural and recombinant rat IL-3
Test Principle
This rat IL-3 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for IL-3 has been precoated onto 96-well plates. Standards(E.coli, I27-C169) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for IL-3 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the rat IL-3 amount of sample captured in plate.
Kit Components
96-well plate precoated with rat IL-3 antibody, 1 Lyophilized recombinant rat IL-3 standard, 10ng/tube×2 Biotinylated rat IL-3 antibody, 130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100) Sample diluent buffer, 30ml Antibody diluent buffer, 12ml ABC diluent buffer, 12ml TMB color developing agent, 10ml TMB stop solution, 10ml
Storage and Stability
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
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