Description
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.
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
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. 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.
3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
4. An isotype control should be used at the same concentration as the antibody of interest.
5. 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.
6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
