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BV421 Mouse Anti-Human CD206 (MMR)
BV421 Mouse Anti-Human CD206 (MMR)

Two-color flow cytometric analysis of CD206 (MMR) expression by GM-CSF-stimulated monocytes. Human peripheral blood mononuclear cells were cultured for 3 days with Recombinant Human GM-CSF (Cat. No. 550068). The cells were harvested, preincubated with Human BD Fc Block™ (Cat. No. 564219/564220), and stained with FITC Mouse Anti-Human CD11c antibody (Cat. No. 561355) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or with BD Horizon BV421 Mouse Anti-Human CD206 (MMR) antibody (Cat. No. 566885; Right Plot). The flow cytometric contour plot showing the correlated expression of CD206 (MMR) [or Ig Isotype control staining] versus CD11c was derived from gated events with the forward and side light scatter characteristics of viable GM-CSF-activated monocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.

Two-color flow cytometric analysis of CD206 (MMR) expression by GM-CSF-stimulated monocytes. Human peripheral blood mononuclear cells were cultured for 3 days with Recombinant Human GM-CSF (Cat. No. 550068). The cells were harvested, preincubated with Human BD Fc Block™ (Cat. No. 564219/564220), and stained with FITC Mouse Anti-Human CD11c antibody (Cat. No. 561355) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or with BD Horizon BV421 Mouse Anti-Human CD206 (MMR) antibody (Cat. No. 566885; Right Plot). The flow cytometric contour plot showing the correlated expression of CD206 (MMR) [or Ig Isotype control staining] versus CD11c was derived from gated events with the forward and side light scatter characteristics of viable GM-CSF-activated monocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.

Product Details
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BD Horizon™
CLEC13D; MMR; macrophage mannose receptor 1
Human (QC Testing)
Mouse BALB/c IgG1, κ
Purified Human MMR
Flow cytometry (Routinely Tested)
5 µl
VII 70802
AB_2869936
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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.

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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant™ Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant™ Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant™ Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant™ Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant™ Stain Buffer Plus (Cat. No. 566385).

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. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  11. Pacific Blue™ is a trademark of Life Technologies Corporation.
566885 Rev. 2
Antibody Details
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15-2

The 15-2 monoclonal antibody specifically recognizes CD206 which is also known as Macrophage mannose receptor (MMR) or C-type lectin domain family 13 member D (CLEC13D). CD206 (MMR) is a ~175 kDa type I transmembrane glycoprotein that is encoded by MRC1 (Mannose receptor C-type 1) which belongs to the mannose receptor family. The extracellular region of CD206 (MMR) is comprised of an N-terminal cysteine-rich domain, followed by a fibronectin type II domain, eight carbohydrate recognition domains (CRD), a transmembrane segment, and a short cytoplasmic tail. CD206 (MMR) is expressed on human macrophages, immature dendritic cells, and endothelial cells. It is not detected on resting monocytes. CD206 (MMR) can function as a Pattern Recognition Receptor (PRR) since it binds to glycoconjugates containing mannose, fucose, or N-acetylglucosamine that are present on the surface of many microorganisms. This receptor enables macrophages and dendritic cells to bind and internalize microbes and their products through endocytosis and phagocytosis and to participate in innate immunity as well as antigen processing related to antigen presentation for adaptive immune responses.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon™ BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon™ BV421, Pacific Blue™, and BD Horizon™ V450 cannot be used simultaneously.

566885 Rev. 2
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
566885 Rev.2
Citations & References
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Development References (7)

  1. Barrett-Bergshoeff M, Noorman F, Bos R, Rijken DC. Monoclonal antibodies against the human mannose receptor that inhibit the binding of tissue-type plasminogen activator.. Thromb Haemost. 1997; 77(4):718-24. (Immunogen: ELISA, Functional assay, Immunohistochemistry, Western blot). View Reference
  2. Kato M, MacDonald K, Munster D, Clark GJ, Hart DNJ. CD206 Macrophage Mannose Receptor Workshop Panel report. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:303-306.
  3. Kato M, Neil TK, Fearnley DB, McLellan AD, Vuckovic S, Hart DN. Expression of multilectin receptors and comparative FITC-dextran uptake by human dendritic cells. Int Immunol. 2000; 12(11):1511-1519. (Clone-specific: Flow cytometry). View Reference
  4. Noorman F, Braat EA, Barrett-Bergshoeff M, et al. Monoclonal antibodies against the human mannose receptor as a specific marker in flow cytometry and immunohistochemistry for macrophages.. J Leukoc Biol. 1997; 61(1):63-72. (Clone-specific: Flow cytometry, Immunohistochemistry, Western blot). View Reference
  5. Nunez R, Filgueira L. Flow cytometric assessment of the reactivity of a panel of monoclonal antibodies (mAb) against two populations of human dendritic cells (DC).. BMC Immunol. 2001; 2:6. (Clone-specific: Flow cytometry). View Reference
  6. Stahl PD, Ezekowitz RA. The mannose receptor is a pattern recognition receptor involved in host defense. Curr Opin Immunol. 1998; 10(1):50-55. (Biology). View Reference
  7. Zola H, Swart B, Boumsell L, Mason DY. Human Leucocyte Differentiation Antigen nomenclature: update on CD nomenclature. Report of IUIS/WHO Subcommittee.. J Immunol Methods. 2003; 275(1-2):1-8. (Clone-specific: Flow cytometry). View Reference
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566885 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.