Preparation And Storage
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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794 or 566349).
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
- This antibody was developed for use in flow cytometry.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
- Cy is a trademark of GE Healthcare.
Companion Products
The TLD-3A12 antibody specifically recognizes CD31, also known as PECAM-1 (platelet endothelial cell adhesion molecule). CD31 is a ~130 kDa integral membrane glycoprotein, a member of the immunoglobulin superfamily, that mediates homophilic and heterophilic cell-cell adhesion. CD31 is expressed on endothelial cells, platelets, and subsets of leucocytes. In the human and mouse, multiple alternatively spliced isoforms have been identified, and this alternative splicing may be involved in the regulation of ligand specificity. CD31-mediated endothelial cell-cell interactions are involved in angiogenesis in the mouse and rat. In addition, PECAM-1 has been demonstrated to play an important role in extravasation of leucocytes in vivo, but in vivo treatment with TLD-3A12 mAb had no discernable effect upon the inflammatory infiltrate in experimental allergic encephalomyelitis. The TLD-3A12 mAb partially blocks the proliferative response of antigen-specific CD4+ T cells to antigen-presenting cells and relevant antigen. This mouse anti-rat CD31 antibody crossreacts with pig endothelial cells and platelets, as determined by immunohistochemical staining of acetone-fixed frozen sections and immunofluorescent staining with flow cytometric analysis, respectively. A suspension of Lewis rat microglial cells was used as the the immunogen.
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.
Development References (5)
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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. (Biology). View Reference
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DeLisser HM, Newman PJ, Albelda SM. Molecular and functional aspects of PECAM-1/CD31. Immunol Today. 1994; 15(10):490-495. (Biology). View Reference
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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
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Wakelin MW, Sanz MJ, Dewar A, et al. An anti-platelet-endothelial cell adhesion molecule-1 antibody inhibits leukocyte extravasation from mesenteric microvessels in vivo by blocking the passage through the basement membrane. J Exp Med. 1996; 184(1):229-239. (Biology). View Reference
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Williams KC, Zhao RW, Ueno K, Hickey WF. PECAM-1 (CD31) expression in the central nervous system and its role in experimental allergic encephalomyelitis in the rat. J Neurosci Res. 1996; 45(6):747-757. (Immunogen: Immunohistochemistry). View Reference
Please refer to Support Documents for Quality Certificates
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.