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PE-CF594 Mouse Anti-Human IL-8
PE-CF594 Mouse Anti-Human IL-8

Two-color flow cytometric analysis of IL-8 expression in stimulated human peripheral blood mononuclear cells. Human PBMC were cultured (6 hr) with Recombinant Human IFN-γ (Cat. No. 554617/554616; 10 ng/ml) and lipopolysaccharide (Sigma, Cat. No. L-8274; 1.0 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The PBMC were harvested and washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) and stained with APC Mouse Anti-Human CD14 antibody (Cat. No. 555399/561708/561383). After washing, the cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ PE-CF594 Mouse IgG2b, κ Isotype Control (Cat. No. 562305; Left Panel) or BD Horizon™ PE-CF594 Mouse Anti-Human IL-8 antibody (Cat. No. 563531; Right Panel) using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-8 (or Ig Isotype control staining) versus CD14 for gated events with the forward and side light-scatter characteristics of intact stimulated PBMC. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Two-color flow cytometric analysis of IL-8 expression in stimulated human peripheral blood mononuclear cells. Human PBMC were cultured (6 hr) with Recombinant Human IFN-γ (Cat. No. 554617/554616; 10 ng/ml) and lipopolysaccharide (Sigma, Cat. No. L-8274; 1.0 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The PBMC were harvested and washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) and stained with APC Mouse Anti-Human CD14 antibody (Cat. No. 555399/561708/561383). After washing, the cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ PE-CF594 Mouse IgG2b, κ Isotype Control (Cat. No. 562305; Left Panel) or BD Horizon™ PE-CF594 Mouse Anti-Human IL-8 antibody (Cat. No. 563531; Right Panel) using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-8 (or Ig Isotype control staining) versus CD14 for gated events with the forward and side light-scatter characteristics of intact stimulated PBMC. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

商品详情
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BD Horizon™
IL8; CXCL8; GCP-1; LYNAP; MDNCF; MONAP; NAP-1; emoctakin
Human (QC Testing)
Mouse IgG2b
Recombinant Human IL-8
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2738263
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


准备和存储

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 BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.

商品通知

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. CF™ is a trademark of Biotium, Inc.
  10. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  11. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  12. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
563531 Rev. 1
抗体详情
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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).

This antibody is conjugated to BD Horizon™ PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg 610/20-nm filter).

  

563531 Rev. 1
格式详情
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
563531 Rev.1
报价单和参考
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研发参考 (9)

  1. Car BD, Meloni F, Luisetti M, Semenzato G, Gialdroni-Grassi G, Walz A. Elevated IL-8 and MCP-1 in the bronchoalveolar lavage fluid of patients with idiopathic pulmonary fibrosis and pulmonary sarcoidosis. Am J Respir Crit Care Med. 1994; 149(3 Pt 1):655-659. (Biology). 查看参考
  2. Emadi S, Clay D, Desterke C, et al. IL-8 and its CXCR1 and CXCR2 receptors participate in the control of megakaryocytic proliferation, differentiation, and ploidy in myeloid metaplasia with myelofibrosis. Blood. 2005; 105(2):464-473. (Clone-specific: Flow cytometry, Immunocytochemistry (cytospins), Immunofluorescence). 查看参考
  3. Hebert CA, Luscinskas FW, Kiely JM, et al. Endothelial and leukocyte forms of IL-8. Conversion by thrombin and interactions with neutrophils. J Immunol. 1990; 145(9):3033-3040. (Biology). 查看参考
  4. Larsen CG, Anderson AO, Appella E, Oppenheim JJ, Matsushima K. The neutrophil-activating protein (NAP-1) is also chemotactic for T lymphocytes. Science. 1989; 243(4897):1464-1466. (Biology). 查看参考
  5. Leonard EJ, Skeel A, Yoshimura T, Noer K, Kutvirt S, Van Epps D. Leukocyte specificity and binding of human neutrophil attractant/activation protein-1. J Immunol. 1990; 144(41323):1323-1330. (Biology). 查看参考
  6. Lizasa H, Matsushima K. IL-8. In: Oppenheim JJ, Feldmann M, Durum SK, Hirano T, Vilcek J, Nicola NA, ed. Cytokine Reference : A compendium of cytokines and other mediators of host defense. San Diego: Academic Press; 2001:1061-1067.
  7. Matsushima K, Oppenheim JJ. Interleukin 8 and MCAF: novel inflammatory cytokines inducible by IL 1 and TNF. Cytokine. 1989; 1(1):2-13. (Biology). 查看参考
  8. Miller LS, Sorensen OE, Liu PT, et al. TGF-alpha regulates TLR expression and function on epidermal keratinocytes. J Immunol. 2005; 174(10):6137-6143. (Clone-specific: ELISA). 查看参考
  9. 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). 查看参考
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563531 Rev. 1

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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.