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PE Mouse Anti-Human CD200
PE Mouse Anti-Human CD200

Profile of anti-CD200 (MRC OX-104) on peripheral blood lymphocytes analyzed by flow cytometry

PE Mouse Anti-Human CD200

Multiparameter flow cytometric analysis of CD200 expression on human peripheral blood leucocyte populations. Whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426/562427) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 555749; Left Plots) or PE Mouse Anti-Human CD200 antibody (Cat. No. 552475/561762; Right Plots). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

        Upper Plots: Bivariate pseudocolor density plots showing the correlated expression of CD200 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations.

        Lower Plots: Bivariate pseudocolor density plots showing the correlated expression of CD200 (or Ig Isotype control staining) versus CD3 signals were derived from gated events with the forward and side-light scatter characteristics of intact lymphocytes.

Profile of anti-CD200 (MRC OX-104) on peripheral blood lymphocytes analyzed by flow cytometry

Multiparameter flow cytometric analysis of CD200 expression on human peripheral blood leucocyte populations. Whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426/562427) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 555749; Left Plots) or PE Mouse Anti-Human CD200 antibody (Cat. No. 552475/561762; Right Plots). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

        Upper Plots: Bivariate pseudocolor density plots showing the correlated expression of CD200 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations.

        Lower Plots: Bivariate pseudocolor density plots showing the correlated expression of CD200 (or Ig Isotype control staining) versus CD3 signals were derived from gated events with the forward and side-light scatter characteristics of intact lymphocytes.

Product Details
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BD Pharmingen™
OX-2; MOX1; MOX2 ; My033
Human (QC Testing)
Mouse IgG1, κ
Human CD200 Recombinant Protein
Flow cytometry (Routinely Tested)
20 µl
VII 70655; IX 40
4345
AB_394398
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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.

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
552475 Rev. 5
Antibody Details
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MRC OX-104

The MRC OX-104 monoclonal antibody specifically binds to CD200. CD200 is a 40-45 kDa type 1 transmembrane glycoprotein and is also referred to as OX2. CD200 is a member of the immunoglobulin superfamily of proteins. It contains two Ig domains, a single transmembrane region and a short cytoplasmic tail. CD200 is expressed on some dendritic cell subsets, and resting and activated T- and B-cells, but not on NK cells, monocytes, granulocytes, or platelets. It has been found on a subset of CD34+ progenitor cells. Interaction of CD200 with its receptor on macrophages induces a downregulation of macrophage activity in a variety of tissues, suggesting a regulatory function.

552475 Rev. 5
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
552475 Rev.5
Citations & References
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Development References (7)

  1. Barclay AN, Wright GJ, Brown MH. CD200 (OX2) Summary and Workshop report: The lymphoid/ neuronal OX2 glycoprotein (CD200) interacts with a novel receptor on macrophages. 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; 2002:471-473.
  2. Cabezon R, Sintes J, Llinas L, Benitez-Ribas D. Analysis of HLDA9 mAbs on plasmacytoid dendritic cell. Immunol Lett. 2011; 134(2):167-173. (Clone-specific: Flow cytometry). View Reference
  3. Ding Y, Ju X, Azlan M, Hart DN, Clark GJ. Screening of the HLDA9 panel on peripheral blood dendritic cell populations.. Immunol Lett. 2011; 134(2):161-6. (Clone-specific: Flow cytometry). View Reference
  4. Hoek RM, Ruuls SR, Murphy CA, et al. Down-regulation of the macrophage lineage through interaction with OX2 (CD200).. Science. 2000; 290(5497):1768-71. (Biology). View Reference
  5. 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.
  6. Wright GJ, Jones M, Puklavec MJ, Brown MH, Barclay AN. The unusual distribution of the neuronal/lymphoid cell surface CD200 (OX2) glycoprotein is conserved in humans. Immunology. 2001; 102(2):173-179. (Biology). View Reference
  7. Wright GJ, Puklavec MJ, Willis AC, et al. Lymphoid/neuronal cell surface OX2 glycoprotein recognizes a novel receptor on macrophages implicated in the control of their function. Immunity. 2000; 13(2):233-242. (Biology). View Reference
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552475 Rev. 5

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