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Immunohistochemical staining for CD62E. Frozen tongue sections of a lipopolysaccharide treated C57BL/6 mouse was reacted with the 10E9.6 antibody. Endothelial cells of the blood vessels express CD62E and can be identifed by the brown staining.
BD Pharmingen™ Purified 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
Immunohistochemistry: The 10E9.6 antibody (Cat. No. 550290) is recommended to test for immunohistochemical staining of acetone-fixed frozen sections. Tissue tested was tongue from LPS treated C57BL/6. The antibody stains endothelial cells in the mouse. The isotype control recommended for use with this antibody is purified rat IgG2a (Cat. No. 559073). For optimal indirect immunohistochemical staining, the 10E9.6 antibody should be titrated (1:10 to 1:50 dilution) and visualized via a three-step staining procedure in combination with biotinylated polyclonal anti-rat Igs (multiple adsorbed) (Cat. No. 559286) as the secondary antibody and Streptavidin-HRP (Cat. No. 550946) together with the DAB detection system (Cat. No. 550880). The clone 10E9.6 is not recommended for formalin-fixed paraffin embedded sections.
A detailed protocol of the immunohistochemical procedure is available at our website, http://www.bdbiosciences.com/support/resources
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
- This antibody has been developed for the immunohistochemistry application. However, a routine immunohistochemistry test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
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.