The G265-8 monoclonal antibody specifically binds to both the 72 and 77 amino acid isoforms of human Interleukin-8 (IL-8). IL-8 is secreted as an 8-9 kDa, non-glycosylated proinflammatory chemokine protein also known as chemokine (C-X-C motif) ligand 8 (CXCL8). IL-8 is synthesized as a 99 amino acid precursor that is proteolytically processed into several isoforms. The 72 amino acid isoform is produced by monocytes, macrophages, granulocytes, epithelial cells, and fibroblasts in response to pro-inflammatory stimuli including cytokines and microbial agents. It is also expressed by endothelial cells, fibroblasts, keratinocytes, lymphocytes, and a variety of tumor cells. In response to IL-4, IL-10 and TGFβ, the cellular production of IL-8 is inhibited. IL-8 is crucial for the activation and recruitment of neutrophils to inflammatory sites. IL-8 is also a chemoattractant for basophils and T-lymphocytes. IL-8 possesses angiogenic activity and can be associated with tumor angiogenesis and metastasis. The 77 amino acid IL-8 isoform is primarily produced by endothelial cells. This larger isoform is reportedly a less potent neutrophil activator than the 72 amino acid isoform. IL-8 binds to and signals through two G-protein-coupled receptors, IL-8RA (CXCR1/CD181) and IL-8RB (CXCR2/CD182).
1. ELISA Detection: For a sandwich ELISA to measure human IL-8 protein levels, the Biotin Mouse Anti-Human IL-8 (Cat. No. 554718) can be used as detector in conjunction with the Purified Mouse Anti-Human IL-8 (Cat. No. 554716) as the capture antibody, with recombinant human IL-8 (Cat. No. 554609) as the standard. The Biotin Mouse Anti-Human IL-8 antibody should be titrated from 0.5-2 µg/ml to determine the optimal concentration for ELISA detection. To obtain standard curves, doubling dilutions of human IL-8 protein ranging from ~1,000 to 1 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our protocols under "ELISA and ELISPOT" at our website: http://www.bdbiosciences.com/us/s/resources. Note: This ELISA pair shows no cross-reactivity with other cytokines or chemokines (e.g., human IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, Il-10, IL-11, IL-12, IL-13, Il-15, GM-CSF, IFN-γ, TNF, LT-α, MCP-1, MCP-2, MIP-1α, MIP-1β, SCF, lymphotactin, VEGF, G-CSF, NT-3, PDGF, CD23, IP-10, GRO, NAP-2, PF4). Note: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assaying serum or plasma samples. For measuring IL-8 in serum or plasma our human IL-8 BD™ OptEIA Set (Cat. No. 555244) or BD OptEIA Kit (Cat. No. 550999) are specifically formulated and recommended. 2. Immunofluorescent Staining and Flow Cytometric Analysis: The G265-8 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate human IL-8 producing cells within mixed cell populations. The PE Mouse Anti-Human IL-8 (Cat. No. 554720) is suitable for these experiments. For specific methodology, please visit our protocols under "Intracellular Flow" at our website: http://www.bdbiosciences.com/us/s/resources.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
