Sandwich ELISA quantitative detection kit for Human IL-16 in cell culture supernates, serum and plasma (heparin, EDTA). Interleukin 16 (IL-16) is a cytokine that released by a variety of cells (including lymphocytes and some epithelial cells) that has been characterized as a chemoattractant for certain immune cells expressing the cell surface molecule CD4. By Southern blot analysis and PCR using a human/rodent somatic cell hybrid mapping panel, The human IL16 is encoded by a single-copy gene on chromosome 15. Using a combination of STS-content mapping, radiation-hybrid mapping, and genetic mapping, it was refined the assignment to 15q26.1. The mouse Il16 gene was mapped to chromosome 7 in a region showing homology of synteny to human 15q26.1. IL-16 was originally described as a factor that could attract activated T cells in humans, it was previously called lymphocyte chemoattractant factor (LCF), and the augmentation of IL16stimulation by CCR5 plays a role in regulation of Th1 cell recruitment and activation at sites of inflammation. Uniprot ID: Q14005 Entrez Gene ID: 3603 Intended Use: This kit is intended for the quantitative determination of human IL-16 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Principle of the Assay: The Interleukin-16 (IL-16) BioAssay(TM) ELISA Kit (Human) is based on standard sandwich enzyme-linked immunosorbent assay technology. A mouse monoclonal antibody specific for IL-16 has been pre-coated onto 96-well plates. Standards (E. coli-expressed recombinant fragment corresponding to P2 thru S130 of human IL-16) and test samples are added to the wells, incubated and then biotinylated detection goat polyclonal antibody specific for IL-16 is added, followed by an incubation step and a washing step with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with PBS or TBS buffer. HRP substrate TMB is used to visualize the HRP-mediated reaction. TMB is catalyzed by HRP to produce a blue colored product that changes into yellow upon addition of acidic stop solution. The intensity of the yellow color is proportional to the amount of human IL-16 in the sample. Detection Range: 31.2pg/ml - 2000pg/ml Sensitivity: <10 pg/ml Precision: Intra-Assay CV: <10% Inter-Assay CV: <10% Cross-reactivity: There is no detectable cross-reactivity with other relevant proteins. Kit Components: 143898A: Microtiter Plate, 1x96wells. Pre-coated with anti-human IL-16 antibody 143898B:IL-16 standard, 2x10ng 143898C: Anti-human IL-16 (Biotin), 1x130ul (dilute 1:100) 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilute 1:100) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml 143702: Wash Buffer, 1x1 packet. PBS powder for preparation of 1L buffer. Storage and Stability: Unopened kit is stable at 4C for 6 months. Assay Summary: 1. Add samples and standards and incubate the plate at 37C for 90 min. Do not wash. 2. Add biotinylated antibody and incubate the plate at 37C for 60 min. Wash plate 3 times with wash buffer. 3. Add ABC working solution and incubate the plate at 37C for 30min. Wash plate 5 times with wash buffer. 4. Add TMB color developing agent and incubate the plate at 37C in dark for 20-25 min. 5. Add Stop solution and read absorbance at 450nm.
