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.
Sensitivity:<10pg/ml
Range:7.8-500pg/ml
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, 1Lyophilized recombinant rat IL-3 standard, 10ng/tube×2Biotinylated rat IL-3 antibody, 130ul (dilution 1:100)Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100)Sample diluent buffer, 30mlAntibody diluent buffer, 12mlABC diluent buffer, 12mlTMB color developing agent, 10mlTMB 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.
