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Biotin Mouse Anti-Human IL-8
Biotin Mouse Anti-Human IL-8

Human IL-8 sandwich ELISA standard curves.  The curves were generated by sandwich ELISAs using Purified Mouse Anti-Human IL-8 (Cat. No. 554716) as capture antibody, doubling dilutions of recombinant IL-8 standards, and Biotin Mouse Anti-Human IL-8 (Cat. No. 554718) as detection antibody. Streptavidin-HRP (Cat. No. 554066) plus TMB substrate (Cat. No. 555214) were used to develop the ELISA. Note that several distinct N-termini exist in IL-8; standard curves were generated with recombinant 72 amino acid (open square curve) or 77 amino acid (filled circle curve) IL-8 standards.

Human IL-8 sandwich ELISA standard curves.  The curves were generated by sandwich ELISAs using Purified Mouse Anti-Human IL-8 (Cat. No. 554716) as capture antibody, doubling dilutions of recombinant IL-8 standards, and Biotin Mouse Anti-Human IL-8 (Cat. No. 554718) as detection antibody. Streptavidin-HRP (Cat. No. 554066) plus TMB substrate (Cat. No. 555214) were used to develop the ELISA. Note that several distinct N-termini exist in IL-8; standard curves were generated with recombinant 72 amino acid (open square curve) or 77 amino acid (filled circle curve) IL-8 standards.

Product Details
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BD Pharmingen™
IL8; CXCL8; GCP-1; LYNAP; MDNCF; MONAP; NAP-1; emoctakin
Human (QC Testing)
Mouse IgG2b
Recombinant Human IL-8
ELISA Detection (Routinely Tested)
0.5 mg/ml
AB_395527
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

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.

Recommended Assay Procedures

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.

Product Notices

  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.
554718 Rev. 5
Antibody Details
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G265-8

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).

554718 Rev. 5
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
554718 Rev.5
Citations & References
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Development References (1)

  1. Matsushima K, Oppenheim JJ. Interleukin 8 and MCAF: novel inflammatory cytokines inducible by IL 1 and TNF. Cytokine. 1989; 1(1):2-13. (Clone-specific). View Reference
554718 Rev. 5

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.