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BV421 Mouse Anti-Human MIP-1β
BV421 Mouse Anti-Human MIP-1β

Flow cytometric analysis of MIP-1β expressed in human peripheral blood mononuclear cells (PBMC). Human PBMC were either unstimulated (Left Panel) or stimulated (Right Panel) 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 2 μM 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 with either a BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or with the BD Horizon™ BV421 Mouse Anti-Human MIP-1β antibody (Cat. No. 562900; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable monocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of MIP-1β expressed in human peripheral blood mononuclear cells (PBMC). Human PBMC were either unstimulated (Left Panel) or stimulated (Right Panel) 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 2 μM 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 with either a BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or with the BD Horizon™ BV421 Mouse Anti-Human MIP-1β antibody (Cat. No. 562900; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable monocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
Macrophage inflammatory protein 1-beta; CCL4; C-C motif chemokine 4; LAG-1
Human (QC Testing)
Mouse IgG1, κ
Recombinant Human MIP-1β
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2737877
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

  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. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  8. Brilliant Violet™ 421 is a trademark of Sirigen.
562900 Rev. 1
Antibody Details
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D21-1351

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 Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

562900 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
562900 Rev.1
Citations & References
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Development References (4)

  1. 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
  2. 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
  3. 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
  4. 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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562900 Rev. 1

Please refer to Support Documents for Quality Certificates


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.