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BB700 Mouse Anti-Human CD31
BB700 Mouse Anti-Human CD31
Multiparameter flow cytometric analysis of CD31 expression on human peripheral blood leucocyte populations. Whole blood was stained with either BD Horizon™ BB700 Mouse IgG1, κ Isotype Control (Cat. No. 566404; Left Plot) or BD Horizon BB700 Mouse Anti-Human CD31 antibody (Cat. No. 566563/566564; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric dot plots showing the correlated expression of CD31 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter signals of intact leucocyte populations. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System.
Multiparameter flow cytometric analysis of CD31 expression on human peripheral blood leucocyte populations. Whole blood was stained with either BD Horizon™ BB700 Mouse IgG1, κ Isotype Control (Cat. No. 566404; Left Plot) or BD Horizon BB700 Mouse Anti-Human CD31 antibody (Cat. No. 566563/566564; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric dot plots showing the correlated expression of CD31 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter signals of intact leucocyte populations. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System.
Product Details
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BD Horizon™
EndoCAM; GPIIA'; PECA1; PECAM1; Platelet endothelial cell adhesion molecule
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl
V P025
5175
AB_2744362
Aqueous buffered solution containing ≤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™ BB700 under optimum conditions, and unconjugated antibody and free BD Horizon BB700 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet for the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. 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.
  6. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
  7. Cy is a trademark of GE Healthcare.
  8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566563 Rev. 1
Antibody Details
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WM59

The WM59 monoclonal antibody specifically binds to platelet endothelial cell adhesion molecule-1, (PECAM-1, PECAM1), which is also known as GPIIA', or EndoCAM. CD31 is a 130 kDa type I transmembrane glycoprotein that belongs to the Ig gene superfamily.  CD31 has wide tissue distribution and is expressed on platelets, monocytes, granulocytes, NK cells, T cell subsets, and in high amounts on endothelial cells. CD31 functions as a vascular endothelial cell adhesion molecule and is involved in the transendothelial migration of leucocytes in inflammatory responses. It might be involved in thrombosis, angiogenesis, and wound healing. The WM59 appears to recognize an epitope proximal to extracellular domain 2 of CD31.

Clone WM59 also cross-reacts with peripheral blood platelets and leukocytes of baboon, and both rhesus and cynomolgus macaque monkeys. The staining intensity of WM59+ platelets is similiar to that observed with peripheral blood platelets from normal human donors. Lymphocytes, monocytes, and granulocytes react less with WM59 than normal human leukocytes.

The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes.   It is a polymer-based tandem dye developed exclusively by BD Biosciences.  With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

566563 Rev. 1
Format Details
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BB700
The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB700
Blue 488 nm
476 nm
695 nm
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Citations & References
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Development References (7)

  1. DeLisser HM, Newman PJ, Albelda SM. Platelet endothelial cell adhesion molecule (CD31). Curr Top Microbiol Immunol. 1993; 184:37-45. (Biology). View Reference
  2. Fawcett J, Buckley C, Holness CL, et al. Mapping the homotypic binding sites in CD31 and the role of CD31 adhesion in the formation of interendothelial cell contacts. J Cell Biol. 1995; 128(6):1229-1241. (Clone-specific: Blocking, ELISA). View Reference
  3. Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. 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; 2002:519-523.
  4. Muller WA, Weigl SA, Deng X, Phillips DM. PECAM-1 is required for transendothelial migration of leukocytes. J Exp Med. 1993; 178(2):449-460. (Biology). View Reference
  5. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
  6. Uguccioni M, Willimann K. Cytokine/Chemokine Receptors: Section 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; 2002:237-243.
  7. Vaporciyan AA, DeLisser HM, Yan HC, et al. Involvement of platelet-endothelial cell adhesion molecule-1 in neutrophil recruitment in vivo.. Science. 1993; 262(5139):1580-2. (Biology). View Reference
View All (7) View Less
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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.