Description
Interleukin-8 (IL-8) is an 8 kD protein containing 72 amino acid residues. It promotes neutrophil chemotaxis and degranulation. The 72 amino acid IL-8 is the predominant form secreted by monocytes and lymphocytes.
Formulation and Purity
Purified human IL-8 was found to be > 98% pure as measured by SDS-PAGE analysis and an absorbance assay based on the Beers-Lambert law. It possesses ≤ 0.1 ng endotoxin per µg human IL-8 as measured using a chromogenic LAL assay. Recombinant human IL-8 is a lyophilized aqueous buffered solution containing 50 µg bovine serum albumin per 1µg recombinant human IL-8. The product contains no preservatives.
Preparation And Storage
Store product at -80°C prior to reconstitution, or for long term storage of stock solutions.The product can be reconstituted in sterile neutral buffer containing carrier protein* such as human or bovine serum albumin, aliquoted and stored at -80°C at a minimum concentration of 25ug/ml for six months. For in vitro biological assay use, we recommend carrier-protein concentrations of 0.5 -1 mg/ml. For use as an ELISA standard we recommend carrier-protein concentrations of 5 - 10 mg/ml. NOTE: Failure to add carrier protein or store at indicated temperatures may result in a loss of activity.
Recommended Assay Procedures
1. ELISA Standard: Recombinant human IL-8 is useful as a quantitative standard for measuring human IL-8 protein levels in an IL-8 specific sandwich ELISA with the purified G265-5 antibody (Cat. No. 554716) as a capture antibody and the biotinylated G265-8 (Cat. No. 554718) as the detection antibody. To obtain linear standard curves, doubling dilutions of this human IL-8 standard from 500-4 pg/ml should be included in each ELISA plate. For specific methodology, please visit the protocols section or chapter on ELISA in the Immune Function Handbook, both of which are posted on our website, www.bdbiosciences.com.
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 human IL-8 in serum or plasma the BD OptEIA™ human IL-8 ELISA Set (Cat. No. 555244) and OptEIA™ human IL-8 ELISA Kit (Cat. No. 550999) are specially formulated and recommended.
2. Representative Biological Activity: Measured Calcium flux using lysed whole blood. The ED50 for these effects is typically 0.15 - 0.30 µg/ml or 0.1 -0.5 ng/ml respectively. Please note that this application is not routinely tested at BD Biosciences.
3. Ligand Blocking Control for Immunofluorescent Staining of Cytokines: Recombinant human IL-8 can be used as a blocking control to demonstrate specificity of IL-8 staining by the directly conjugated G265-8 antibody (Cat. No. 554720). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. For specific methodology, please visit please visit the protocols section or chapter on intracellular staining in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com.
*Note: Carrier proteins should be pre-screened for possible effects in an appropriate experimental system. Carrier proteins may effect experimental results due to toxicity, high endotoxin levels or possible blocking activity.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- IL-8 is protected under U.S. Patent Nos. 5,652,338 and 5,698,196.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Development References (3)
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Bauser R, Proudfoot AE. Calcium Mobilization. In: Proudfoot AE, Wells TN, Power CA, ed. Chemokine Protocols, Volume 138. Totowa, New Jersey: Humana Press; 2000:143-148.
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Matsushima K, Oppenheim JJ. Interleukin 8 and MCAF: novel inflammatory cytokines inducible by IL 1 and TNF. Cytokine. 1989; 1(1):2-13. (Biology). View Reference
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Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: Blocking). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.
