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Flow cytometric analysis of MIP-1β expressed in human peripheral blood mononuclear cells (PBMC). Human PBMC were either unstimulated (Left Plot) or stimulated (Right Plot) with 20 ng/mL Recombinant Human IFN-γ (Cat. No. 554616) for one hour followed by overnight incubation with 1 μg/mL LPS (Sigma-Aldrich, Cat. No. L-8272) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724). The PBMC were harvested, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histograms) or with BD Horizon™ R718 Mouse Anti-Human MIP-1β antibody (Cat. No. 567090/567249; solid line histograms). The fluorescence histograms showing the expressed levels of MIP-1β (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
BD Horizon™ R718 Mouse Anti-Human MIP-1β
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD™ CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD™ CompBeada to ensure that BD™ CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. 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.
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
Companion Products
The D21-1351 monoclonal antibody specifically binds to the human CC chemokine, MIP-1β (macrophage inflammatory protein-1β). Human MIP-1β shares approximately 75% homology with mouse MIP-1β at the amino acid level. Expression of MIP-1β in human peripheral blood cells is induced by proinflammatory and mitogenic stimuli. MIP-1β is a chemoattractant for monocytes and lymphocytes. Human MIP-1β binds to receptors, CCR5 and CCR8. The human MIP-1β gene has been mapped to chromosome 17q11. The immunogen used to generate D21-1351 hybridoma was recombinant human MIP-1β.
The antibody was conjugated to BD Horizonx™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.
Development References (8)
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Bernardini G, Hedrick J, Sozzani S. Identification of the CC chemokines TARC and macrophage inflammatory protein-1 beta as novel functional ligands for the CCR8 receptor. J Immunol. 1998; 28(2):582-588. (Biology). View Reference
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Combadiere C, Ahuja SK, Tiffany HL, Murphy PM. Cloning and functional expression of CC CKR5, a human monocyte CC chemokine receptor selective for MIP-1(alpha), MIP-1(beta), and RANTES. J Leukoc Biol. 1996; 60(1):147-152. (Biology). View Reference
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Kiniry BE, Hunt PW, Hecht FM, Somsouk M, Deeks SG, Shacklett BL. Differential Expression of CD8(+) T Cell Cytotoxic Effector Molecules in Blood and Gastrointestinal Mucosa in HIV-1 Infection. J Immunol. 2018; 200(5):1876-1888. (Clone-specific: Flow cytometry). View Reference
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Lu LL, Smith MT, Yu KKQ, et al. IFN-γ-independent immune markers of Mycobacterium tuberculosis exposure.. Nat Med. 2019; 25(6):977-987. (Clone-specific: Flow cytometry). View Reference
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Lu LL, Smith MT, Yu KKQ, et al. IFN-γ-independent immune markers of Mycobacterium tuberculosis exposure.. Nat Med. 2019; 25(6):977-987. (Clone-specific: Flow cytometry). View Reference
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Peper JK, Bösmüller HC, Schuster H, et al. HLA ligandomics identifies histone deacetylase 1 as target for ovarian cancer immunotherapy.. Oncoimmunology. 2016; 5(5):e1065369. (Clone-specific: Flow cytometry). 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: Flow cytometry, IC/FCM Block). View Reference
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Raport CJ, Gosling J, Schweickart VL, Gray PW, Charo IF. Molecular cloning and functional characterization of a novel human CC chemokine receptor (CCR5) for RANTES, MIP-1beta, and MIP-1alpha. J Biol Chem. 1996; 271(29):17161-17166. (Biology). View Reference
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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.