Figure 1. PBMC supernatants, untreated (-) or PHA-treated (10 µg/ml; +) were removed after 1 and 5 d. hIL-8 levels in the supernatants were quantified using SignalKine™ Human IL-8 Sandwich ELISA Kit.
Typical Data. Human IL-8 Standard Curves, one diluted in SignalKine™ Sample Diluent S01, a second diluted in
SignalKine™ Sample Diluent S02 and a third in cell culture media (RPMI + 10% FBS). Although these are typical of the standard curves that will be generated using this kit, a new set of standards should be run for each new experiment.
SignalKine™ Human IL-8 Sandwich ELISA Kit from Cell Signaling Technology (CST) is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects human IL-8 (hIL-8) in multiple matrices. An hIL-8 Rabbit mAb has been coated onto low volume microwells. hIL-8 standards and hIL-8-containing samples are added to the wells and captured by the coated antibody. Following a washing step, a biotinylated hIL-8 Detection Rabbit mAb is added to detect the captured hIL-8. HRP-linked Streptavidin is then used for detection of the biotinylated hIL-8 Detection Rabbit mAb. HRP substrate, TMB, is added for color development. The magnitude of absorbance for this developed color is proportional to the quantity of hIL-8 in the sample.
SignalKine™ Human IL-8 Sandwich ELISA Kit detects hIL-8 in multiple matrices that can be quantified by generating a standard curve with the recombinant hIL-8 protein standard provided. The hIL-8 standard range is from 15.6 to 1000 pg/ml. Samples containing higher levels of hIL-8 can be diluted to fit into the standard range.
IL-8 is the prototypical CXC chemokine. It is best known for effects on neutrophils, specifically its ability to act as a chemoattractant and activate degranulation and respiratory burst (1,2,3). IL-8 is produced by a number of cell types, including monocytes, T cells, neutrophils, fibroblasts, endothelial cells, and others (1). In addition to its effects on neutrophils, IL-8 promotes angiogenesis, inhibits endothelial cell apoptosis, and promotes the proliferation of melanoma cells in an autocrine fashion (1). As a result, IL-8 may be a contributing factor to the development of certain cancers (1,2). There are two distinct G protein-coupled receptors for IL-8, CXCR1 and CXCR2 (1). Ligand binding induces Ca2+ mobilization and activates the PI3K, PKC, Rho, and Rac pathways (1,3).
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