Flow cytometric analysis of MIP-1β on human PBMC. Human PBMC were stimulated with 20 ng/mL 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™ (Cat. No. 554724). Left Panel: The PBMC were harvested, fixed, permeabilized with BD Cytofix/CytoPerm™ Plus Kit (with BD GolgiStop) (Cat. No. 554715), and stained with either a FITC Mouse IgG1, κ isotype control (Cat. No. 555909; shaded) or with the FITC Mouse Anti-Human MIP-1β antibody (Cat. No. 560565; unshaded). Right Panel: Both unstimulated (shaded) and stimulated PBMC (unshaded) were harvested, fixed, permeabilized with BD Cytofix/CytoPerm Plus Kit (with GolgiStop), and stained with the FITC Mouse Anti-Human MIP-1β antibody. Histograms were derived from gated events based on light scattering characteristics for monocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Flow cytometric analysis of MIP-1β on human PBMC. Human PBMC were stimulated with 20 ng/mL 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™ (Cat. No. 554724). Left Panel: The PBMC were harvested, fixed, permeabilized with BD Cytofix/CytoPerm™ Plus Kit (with BD GolgiStop) (Cat. No. 554715), and stained with either a FITC Mouse IgG1, κ isotype control (Cat. No. 555909; shaded) or with the FITC Mouse Anti-Human MIP-1β antibody (Cat. No. 560565; unshaded). Right Panel: Both unstimulated (shaded) and stimulated PBMC (unshaded) were harvested, fixed, permeabilized with BD Cytofix/CytoPerm Plus Kit (with GolgiStop), and stained with the FITC Mouse Anti-Human MIP-1β antibody. Histograms were derived from gated events based on light scattering characteristics for monocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry 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)
20 µl
AB_1645489
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 FITC under optimum conditions, and unreacted FITC was removed.
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation/Permeabilization Solution Kit with BD GolgiStop™ RUO
Size
250 Tests
Cat No.
554715
Protein Transport Inhibitor (Containing Monensin) RUO
Size
0.7 mL
Cat No.
554724
FITC Mouse IgG1, κ Isotype Control RUO
Size
100 Tests
Cat No.
555909
Recombinant Human IFN-γ RUO
Size
25 µg
Cat No.
554616
560565 Rev. 2
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.
560565 Rev. 2
Format Details
FITC
Fluorescein (FITC) is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 518-nm. FITC is designed to be excited by the Blue laser (488-nm) and detected using an optical filter centered near 520 nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
FITC
Blue 488 nm
494 nm
518 nm
560565 Rev.2
Citations & References
View product citations for antibody "560565" on CiteAb
Development References (9)
-
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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Lipes MA, Napolitano M, Jeang KT, Chang NT, Leonard WJ. Identification, cloning, and characterization of an immune activation gene. Proc Natl Acad Sci U S A. 1988; 85(24):9704-9708. (Biology). View Reference
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Napolitano M, Seamon KB, Leonard WJ. Identification of cell surface receptors for the Act-2 cytokine. J Exp Med. 1990; 172(1):285-289. (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
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Samson M, Labbe O, Mollereau C, Vassart G, Parmentier M. Molecular cloning and functional expression of a new human CC-chemokine receptor gene. Biochemistry. 1996; 35(11):3362-3367. (Biology). View Reference
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Sherry B, Tekamp-Olson P, Gallegos C. one of those components, macrophage inflammatory protein 1 beta. J Exp Med. 1988; 168(6):2251-2259. (Biology). View Reference
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Ziegler SF, Tough TW, Franklin TL, Armitage RJ, Alderson MR. Induction of macrophage inflammatory protein-1 beta gene expression in human monocytes by lipopolysaccharide and IL-7. J Immunol. 1991; 147(7):2234-2239. (Biology). View Reference
560565 Rev. 2
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.