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PE Mouse Anti-Human IL-8 G265-8 RUO

PE Mouse Anti-Human IL-8 

Clone  G265-8   (RUO)

  • Brand BD Pharmingen™
  • Alternative Name IL8; CXCL8; GCP-1; LYNAP; MDNCF; MONAP; NAP-1; emoctakin
  • Concentration 0.2 mg/ml
  • Isotype Mouse IgG2b
  • Reactivity Human (QC Testing) 
  • Application Intracellular staining (flow cytometry) (Routinely Tested) 
  • Immunogen Recombinant Human IL-8
  • Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
  • Regulatory Status RUO
Product Details Recommended Assay References

Description

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).

Format
  • Format PE
  • Excitation Source Blue 488 nm,Green 532 nm,Yellow/Green 561 nm
  • Excitation Max 496 nm
  • Emission Max 578 nm

R-phycoerythrin (PE) is an accessory photosynthetic pigment found in red algae. It exists in vitro as a 240-kDa protein with 23 phycoerythrobilin chromophores per molecule. This makes PE one of the brightest fluorochromes for flow cytometry applications, but its photobleaching properties make it unsuitable for fluorescence microscopy.

Other Formats →

Suggested Companion Products

PE Mouse IgG2b, κ Isotype Control RUO
0.1 mg Cat No: 555058

FITC Mouse Anti-Human CD14 M5E2 RUO
100 Tests Cat No: 555397

Recombinant Human IL-8 RUO
20 µg Cat No: 554609

BD Cytofix/Cytoperm Plus Kit (with BD GolgiStop) RUO
250 Tests Cat No: 554715

Protein Transport Inhibitor (Containing Monensin) RUO
0.7 mL Cat No: 554724

Purified Mouse Anti-Human IL-8 G265-8 RUO
0.1 mg Cat No: 554717

Purified Mouse Anti-Human IL-8 G265-8 RUO
0.25 mg Cat No: 550419

Biotin Mouse Anti-Human IL-8 G265-8 RUO
0.5 mg Cat No: 554718

Purified Mouse Anti-Human IL-8 G265-5 RUO
0.5 mg Cat No: 554716

Human IL-8 ELISA Set RUO
20 Plate(s) Cat No: 555244

Human IL-8 ELISA Kit II RUO
2 Plate(s) Cat No: 550999

Resources & Tools
 Spectrum Viewer  Panel Designer  Spectrum Viewer  Download TDS  Regulatory Document Website
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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.


Immunofluorescent Staining and Flow Cytometry: The PE Mouse Anti-Human IL-8 (Cat. No. 554720) can be used for multicolor immunofluorescent staining and flow cytometric analyses to identify and enumerate IL-8 producing cells within mixed cell populations (see figure). For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/million cells). For specific methodology, please visit our protocols under "Intracellular Flow" at our website: http://www.bdbiosciences.com/us/s/resources/.

A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated G265-8 antibody with ligand (e.g., recombinant human IL-8; Cat. No. 554609) prior to staining, or 2) pre-block the fixed/ permeabilized cells with Purified Mouse Anti-Human IL-8 (Cat. No. 554717/550419) prior to staining. The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. An appropriate PE-mouse IgG2b isotype control to use on fixed and permeabilized cells is PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058).

Cytokine ICC: The G265-8 antibody is useful for immunocytochemical staining. Purified Mouse Anti-Human IL-8 (Cat. No. 550419/554717) is tested in the ICC application.

ELISA Detection: The Biotin Mouse Anti-Human IL-8 antibody (Catalog No. 554718) is useful as a detection antibody in a sandwich ELISA for measuring human IL-8 protein levels. Biotin Mouse Anti-Human IL-8 antibody can be paired with the Purified Mouse Anti-Human IL-8 (Cat. No. 554716) with recombinant human IL-8 (Cat. No. 554609) as the standard. This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. For detection of IL-8 in serum or plasma, the Human IL-8 BD OptEIA™ ELISA Set (Cat. No. 555244) or OptEIA™ ELISA Kit (Cat. No. 550999) is recommended.


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

  2. 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. View reference

554720 Rev. 3
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