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 Alexa Fluor® 700 Mouse IgG1, κ Isotype Control (Cat. No. 557882; dashed line histogram) or with the Alexa Fluor® 700 Mouse Anti-Human MIP-1β antibody (Cat. No. 561278; 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.
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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 Alexa Fluor® 700 Mouse IgG1, κ Isotype Control (Cat. No. 557882; dashed line histogram) or with the Alexa Fluor® 700 Mouse Anti-Human MIP-1β antibody (Cat. No. 561278; 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
BD Pharmingen™
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_10612008
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 700 under optimum conditions, and unreacted Alexa Fluor® 700 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Product Notices
- 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 www.bdbiosciences.com/us/s/resources for technical protocols.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
Protein Transport Inhibitor (Containing Monensin) RUO
Size
0.7 mL
Cat No.
554724
Alexa Fluor® 700 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557882
Recombinant Human IFN-γ RUO
Size
25 µg
Cat No.
554616
561278 Rev. 1
Antibody Details
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.
561278 Rev. 1
Format Details
Alexa Fluor™ 700
Alexa Fluor™ 700 dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 697 nm and an emission maximum (Em Max) at 719-nm. Alexa Fluor™ 700 is designed to be excited by the Red (627–640-nm) laser and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 700
Red 627-640 nm
697 nm
719 nm
561278 Rev.1
Citations & References
View product citations for antibody "561278" on CiteAb
Development References (4)
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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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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
561278 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.