The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/1000.
IHC - Wholemount
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 83 kDa).
We recommend using 3% milk as the blocking agent for Western blot.
Application notesIs unsuitable for IHC-Fr or IHC-P.
FunctionInduces susceptibility to atherosclerosis (By similarity). Cell adhesion molecule which is required for leukocyte transendothelial migration (TEM) under most inflammatory conditions. Tyr-690 plays a critical role in TEM and is required for efficient trafficking of PECAM1 to and from the lateral border recycling compartment (LBRC) and is also essential for the LBRC membrane to be targeted around migrating leukocytes. Prevents phagocyte ingestion of closely apposed viable cells by transmitting 'detachment' signals, and changes function on apoptosis, promoting tethering of dying cells to phagocytes (the encounter of a viable cell with a phagocyte via the homophilic interaction of PECAM1 on both cell surfaces leads to the viable cell's active repulsion from the phagocyte. During apoptosis, the inside-out signaling of PECAM1 is somehow disabled so that the apoptotic cell does not actively reject the phagocyte anymore. The lack of this repulsion signal together with the interaction of the eat-me signals and their respective receptors causes the attachment of the apoptotic cell to the phagocyte, thus triggering the process of engulfment). Isoform Delta15 is unable to protect against apoptosis. Modulates BDKRB2 activation. Regulates bradykinin- and hyperosmotic shock-induced ERK1/2 activation in human umbilical cord vein cells (HUVEC).
Tissue specificityExpressed on platelets and leukocytes and is primarily concentrated at the borders between endothelial cells. Isoform Long predominates in all tissues examined. Isoform Delta12 is detected only in trachea. Isoform Delta14-15 is only detected in lung. Isoform Delta14 is detected in all tissues examined with the strongest expression in heart. Isoform Delta15 is expressed in brain, testis, ovary, cell surface of platelets, human umbilical vein endothelial cells (HUVECs), Jurkat T-cell leukemia, human erythroleukemia (HEL) and U937 histiocytic lymphoma cell lines (at protein level).
DomainThe Ig-like C2-type domains 2 and 3 contribute to formation of the complex with BDKRB2 and in regulation of its activity.
Post-translational modificationsPhosphorylated on Ser and Tyr residues after cellular activation. In endothelial cells Fyn mediates mechanical-force (stretch or pull) induced tyrosine phosphorylation.
Cellular localizationMembrane. Cell junction. Localizes to the lateral border recycling compartment (LBRC) and recycles from the LBRC to the junction in resting endothelial cells and Cell junction. Localizes to the lateral border recycling compartment (LBRC) and recycles from the LBRC to the junction in resting endothelial cells.
Immunocytochemistry/ Immunofluorescence - Anti-CD31 antibody [P2B1] (ab24590)This image is courtesy of an Abreview submitted by an anonymous researcher on 7 December 2005.
Positive immunocytochemical staining of human primary choroidal endothelial cells using ab24590 at a dilution of 1/100. The tissue was fixed with paraformaldehyde and the antibody was incubated for 3 hours. An Alexa Fluor® 488 goat anti-mouse antibody was used as a secondary antibody.
Images of isolated capillaries triple labeled for EC marker protein CD31 (red channel), PC marker protein NG-2 (green channel), and Hoechst for nuclei (blue channel).
Cochlear stria vascularis capillaries were isolated from 6-week-old CBA/CaJ mice. The capillary isolation was achieved with a special procedure we developed and named the “sandwich-dissociation” method. The tissue samples were fixed in 4% paraformaldehyde at 4°C for 2 h, washed in PBS for 30 min, permeabilized in 0.5% Triton X-100, then immunoblocked in a solution of 10% goat serum in 1% bovine albumin in PBS for 30 min. The specimens were incubated with primary antibodies and secondary antibodies (goat Alexa Fluo 488-conjugated anti-mouse and anti-rabbit IgG antibodies).
Immunocytochemistry/ Immunofluorescence - Anti-CD31 antibody [P2B1] (ab24590)This image is courtesy of an anonymous Abreview
ab24590 staining CD31 in the HEK 293T cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 1% Triton and blocked with 3% BSA for 1 hour at RT. Samples were incubated with primary antibody (1/50 in PBS + 3% BSA) for 1 hour. A Alexa Fluor® 555-conjugated Donkey anti-mouse polyclonal was used as the secondary antibody (1/500).