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BUV737 Rat Anti-Mouse CD31
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This product is the replacement for [565097].
BUV737 Rat Anti-Mouse CD31

Two-color flow cytometric analysis of CD31 expression on mouse bone marrow cells. Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553089/553090/561878) and either BD Horizon™ BUV737 Rat IgG2a, κ Isotype Control (Cat. No. 612760; Left Plot) or BD Horizon BUV737 Rat Anti-Mouse CD31 antibody (Cat. No. 612802; Right Plot) at 0.5 µg/test. Two-color flow cytometric contour plots showing the correlated expression of CD31 (or Ig Isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Two-color flow cytometric analysis of CD31 expression on mouse bone marrow cells. Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553089/553090/561878) and either BD Horizon™ BUV737 Rat IgG2a, κ Isotype Control (Cat. No. 612760; Left Plot) or BD Horizon BUV737 Rat Anti-Mouse CD31 antibody (Cat. No. 612802; Right Plot) at 0.5 µg/test. Two-color flow cytometric contour plots showing the correlated expression of CD31 (or Ig Isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Horizon™
EndoCAM; GPIIA; PECA1; PECAM1; Platelet endothelial cell adhesion molecule
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2a, κ
129/Sv mouse-derived endothelioma cell line tEnd.1
Flow cytometry (Routinely Tested)
0.2 mg/ml
18613
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 BUV737 under optimum conditions, and unconjugated antibody and free BD Horizon BUV737 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. This will 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 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).

Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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 Ultraviolet 737 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
  6. 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.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
612802 Rev. 2
Antibody Details
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MEC 13.3

The MEC13.3 antibody specifically recognizes CD31, also known as PECAM-1 (Platelet Endothelial Cell Adhesion Molecule-1). CD31 is a 130 kDa integral membrane protein, a member of the immunoglobulin superfamily, that mediates cell-to-cell adhesion. CD31 is expressed constitutively on the surface of adult and embryonic endothelial cells and is also expressed on many peripheral leukocytes and platelets. It has also been detected on bone marrow-derived hematopoietic stem cells and embryonic stem cells. CD31 is involved in the transendothelial emigration of neutrophils, and neutrophil PECAM-1 appears to be down-regulated after extravasation into inflamed tissues. Multiple alternatively spliced isoforms are detected during early post-implantation embryonic development; this alternative splicing is involved in the regulation of ligand specificity. CD38 and vitronectin receptor (αvβ3 integrin, CD51/CD61) are proposed to be ligands for CD31. CD31-mediated endothelial cell-cell interactions are involved in angiogenesis. The MEC13.3 mAb inhibits a variety of in vitro and in vivo functions mediated by CD31.

       The antibody was conjugated to BD Horizon BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 737 nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 nm filter. Due to the excitation of the acceptor dye by the red laser line, there may be significant spillover into red laser detectors with filters in the 700-720 nm range.

612802 Rev. 2
Format Details
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BUV737
The BD Horizon Brilliant™ Ultraviolet 737 (BUV737) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 735-nm. BUV737, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 740-nm (e.g., 740/35 bandpass filter). The acceptor dye can be excited by the Red (628–640nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BUV737
Ultraviolet 355 nm
350 nm
735 nm
612802 Rev.2
Citations & References
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Development References (12)

  1. Baldwin HS, Shen HM, Yan HC, et al. Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31): alternatively spliced, functionally distinct isoforms expressed during mammalian cardiovascular development. Development. 1994; 120(9):2539-2953. (Clone-specific: Blocking, Flow cytometry, Immunohistochemistry). View Reference
  2. Christofidou-Solomidou M, Nakada MT, Williams J, Muller WA, DeLisser HM. Neutrophil platelet endothelial cell adhesion molecule-1 participates in neutrophil recruitment at inflammatory sites and is down-regulated after leukocyte extravasation. J Immunol. 1997; 158(10):4872-4878. (Clone-specific: Flow cytometry, Inhibition, In vivo exacerbation). View Reference
  3. DeLisser HM, Christofidou-Solomidou M, Strieter RM, et al. Involvement of endothelial PECAM-1/CD31 in angiogenesis. Am J Pathol. 1997; 151(3):671-677. (Clone-specific: Inhibition, In vivo exacerbation). View Reference
  4. DeLisser HM, Newman PJ, Albelda SM. Molecular and functional aspects of PECAM-1/CD31. Immunol Today. 1994; 15(10):490-495. (Biology). View Reference
  5. Duncan GS, Andrew DP, Takimoto H, et al. Genetic evidence for functional redundancy of Platelet/Endothelial cell adhesion molecule-1 (PECAM-1): CD31-deficient mice reveal PECAM-1-dependent and PECAM-1-independent functions. J Immunol. 1999; 162(5):3022-3030. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
  6. Famiglietti J, Sun J, DeLisser HM, Albelda SM. Tyrosine residue in exon 14 of the cytoplasmic domain of platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) regulates ligand binding specificity. J Cell Biol. 1997; 138(6):1425-1435. (Biology). View Reference
  7. Horenstein AL, Stockinger H, Imhof BA, Malavasi F. CD38 binding to human myeloid cells is mediated by mouse and human CD31. Biochem J. 1998; 330(3):1129-1135. (Biology). View Reference
  8. Ling V, Luxenberg D, Wang J, et al. Structural identification of the hematopoietic progenitor antigen ER-MP12 as the vascular endothelial adhesion molecule PECAM-1 (CD31). Eur J Immunol. 1997; 27(2):509-514. (Clone-specific: Flow cytometry). View Reference
  9. Piali L, Hammel P, Uherek C, et al. CD31/PECAM-1 is a ligand for alpha v beta 3 integrin involved in adhesion of leukocytes to endothelium. J Cell Biol. 1995; 130(2):451-460. (Biology). View Reference
  10. Rosenblum WI, Murata S, Nelson GH, Werner PK, Ranken R, Harmon RC. Anti-CD31 delays platelet adhesion/aggregation at sites of endothelial injury in mouse cerebral arterioles. Am J Pathol. 1994; 145(1):33-36. (Clone-specific: Flow cytometry, Immunohistochemistry, Immunoprecipitation, In vivo exacerbation). View Reference
  11. Vanzulli S, Gazzaniga S, Braidot MF, et al. Detection of endothelial cells by MEC 13.3 monoclonal antibody in mice mammary tumors. Biocell. 1997; 21(1):39-46. (Clone-specific: Immunohistochemistry). View Reference
  12. Vecchi A, Garlanda C, Lampugnani MG, et al. Monoclonal antibodies specific for endothelial cells of mouse blood vessels. Their application in the identification of adult and embryonic endothelium. Eur J Cell Biol. 1994; 63(2):247-254. (Immunogen: ELISA, Flow cytometry, Fluorescence microscopy, Immunofluorescence, Immunohistochemistry, Immunoprecipitation). View Reference
View All (12) View Less
612802 Rev. 2

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