The monoclonal antibody MEC7.46 recognizes the mouse form of the platelet-endothelial cell adhesion molecule (PECAM)-1 (CD31). PECAM-1 is a member of the immunoglobulin superfamily. This heavily glycosylated protein is found in the entire vascular endothelium of adult mice and functions in mediating cellular adhesion by heterophilic and homophilic mechanisms. PECAM-1 is detected within the lymphopoietic islands in the spleen of newborn (day 12) and in the bone marrow of adult mice. Capillary endothelial cells of adult mice also express PECAM-1.
The reactivity of the monoclonal antibody MEC7.46 is restricted to the isoform of the molecule that is selectively expressed by endothelial cells. The antibody precipitates a 130 kDa molecule present on the membrane of endothelial cells of all mouse blood vessels both in normal, inflamed and tumor tissues. The antigen is predominantly present at the lateral borders of endothelial cells as described for human PECAM-1. Staining of MEC7.46 can be seen on capillaries, veins, arteries and liver sinusoids.
Flow cytometry, Frozen sections, Immuno fluorescence, Immuno precipitation, Paraffin sections
FC: Antibody MEC7.46 stains the extracellular domain of mouse PECAM-1; 106 cells were used per sample. No fixation or permeabilization needed. Epitope is sensitive to trypsin! (Ref.1)
F: Tissue sections were fixed in acetone for 10 min at RT and pretreated with 0.01% hydrogen peroxide to quench endogenous peroxidases. Normal rabbit serum was used as blocking agent. As (Ref.1).
P: Serial paraffin-embedded 4 μm sections were stained by H&E, PAS and Azan Histochemistry. The distribution of endothelial (Mec7.46), matrix (laminin) and macrophage (FA/11) markers was evaluated using a standard three-step ABC method and development in 3-amino-9-ethylcarbazole solution. Sections were counterstained for 45 seconds with Harris' haematoxylin at room temperature and mounted in Imsol-mount medium (ref. 3).
IP: Lysate of 35S-Methionine labeled 5HV cells was used. Molecule of ~130 kDa.
Dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.
Mouse t-end.1 cells
L929 cells (Fibroblasts)