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)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Recombinant Human IL-8
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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 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.
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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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.