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Flow cytometric analysis using BD OptiBuild™ BV421 Rat Anti-Mouse CD62E antibody (Cat. No. 740027; solid line histogram) on bEnd.3 cell line, with Ig Isotype Control (dotted line histogram). Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
BD OptiBuild™ BV421 Rat Anti-Mouse CD62E
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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 (including BD OptiBuild Brilliant reagents) 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).
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 Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
Companion Products
The 10E9.6 monoclonal antibody specifically binds to the 97-110 kDa cell surface glycoprotein E-selectin (CD62E), also known as endothelial-leukocyte adhesion molecule-1 (ELAM-1), which is expressed on endotoxin- or cytokine-stimulated mouse endothelial cells. A suspension of TNFα stimulated mouse brain capillary endothelioma cells, from the cell line bEnd.3, was used as the immunogen. The epitope recognized by mAb 10E9.6 has been mapped to the first and/or second complement regulatory protein repeat domains of E-selectin. The 10E9.6 antibody has been reported to block binding of a monocyte cell line to E-selectin in vitro and to block neutrophil migration in BALB/c, but not C57BL/6 mice. It has no effect on leukocyte rolling in TNFα-treated mouse venules or on in vitro adhesion of myeloid cells to E-selectin. Studies have demonstrated that Cutaneous Lymphocyte Antigen (CLA), recognized by mAb HECA-452 (Cat. no. 555946), may be a ligand for CD62E.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (7)
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Borges E, Pendl G, Eytner R, Steegmaier M, Zollner O, Vestweber D. The binding of T cell-expressed P-selectin glycoprotein ligand-1 to E- and P-selectin is differentially regulated. J Biol Chem. 1997; 272(45):28786-28792. (Biology). View Reference
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Bosse R, Vestweber D. Only simultaneous blocking of the L- and P-selectin completely inhibits neutrophil migration into mouse peritoneum. Eur J Immunol. 1994; 24(12):3019-3024. (Immunogen: Blocking, ELISA, Immunoprecipitation). View Reference
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Eppihimer MJ, Wolitzky B, Anderson DC, Labow MA, Granger DN. Heterogeneity of expression of E- and P-selectins in vivo. Circ Res. 1996; 79(3):560-569. (Biology: Blocking). View Reference
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Ley K, Bullard DC, Arbones ML, et al. Sequential contribution of L- and P-selectin to leukocyte rolling in vivo. J Exp Med. 1995; 181(2):669-675. (Biology). View Reference
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Pendl GG, Robert C, Steinert M, et al. Immature mouse dendritic cells enter inflamed tissue, a process that requires E- and P-selectin, but not P-selectin glycoprotein ligand 1. Blood. 2002; 99(3):946-956. (Biology). View Reference
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Ramos CL, Kunkel EJ, Lawrence MB, et al. Differential effect of E-selectin antibodies on neutrophil rolling and recruitment to inflammatory sites. Blood. 1997; 89(8):3009-3018. (Immunogen: Blocking). View Reference
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Weller A, Isenmann S, Vestweber D. Cloning of the mouse endothelial selectins. Expression of both E- and P-selectin is inducible by tumor necrosis factor alpha. J Biol Chem. 1992; 267(21):15176-15183. (Biology). 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.