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™ 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.
Storage and Stability:Unopened kit is stable at 4°C for 6 months.
Assay Summary:1. Add samples and standards and incubate the plate at 37°C for 90 min. Do not wash. 2. Add biotinylated antibody and incubate the plate at 37°C for 60 min. Wash plate 3 times with wash buffer
3. Add ABC working solution and incubate the plate at 37°C for 30min. Wash plate 5 times with wash buffer
4. Add TMB color developing agent and incubate the plate at 37°C in dark for 20-25 min
5. Add Stop solution and read absorbance at 450nm.
