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BB515 Mouse Anti-Human CD31
BB515 Mouse Anti-Human CD31

Multiparameter flow cytometric analysis of CD31 expression on human peripheral blood leucocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Human whole blood was stained with either BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; Left Panel), BD Horizon BB515 Mouse Anti-Human CD31 antibody (Cat. No. 564630; Middle Panel), or FITC Mouse Anti-Human CD31 antibody (Cat. No. 555445/560984; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202).  Flow cytometric contour plots showing the correlated expression of CD31 (or Ig Isotype control staining) versus side light-scatter signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Multiparameter flow cytometric analysis of CD31 expression on human peripheral blood leucocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Human whole blood was stained with either BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; Left Panel), BD Horizon BB515 Mouse Anti-Human CD31 antibody (Cat. No. 564630; Middle Panel), or FITC Mouse Anti-Human CD31 antibody (Cat. No. 555445/560984; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202).  Flow cytometric contour plots showing the correlated expression of CD31 (or Ig Isotype control staining) versus side light-scatter signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II 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_2738872
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™ BB515 under optimum conditions and unconjugated antibody was removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence 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. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564630 Rev. 3
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 BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

564630 Rev. 3
Format Details
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB515
Blue 488 nm
490 nm
515 nm
564630 Rev.3
Citations & References
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Development References (6)

  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. Fornasa G, Groyer E, Clement M, et al. TCR stimulation drives cleavage and shedding of the ITIM receptor CD31. J Immunol. 2010; 184(10):5485-5492. (Clone-specific: ELISA, Flow cytometry). View Reference
  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. 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 (6) View Less
564630 Rev. 3

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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.