|Detection of Mouse CD31/PECAM‑1 by Western Blot. Western blot shows lysates of bEnd.3 mouse endothelioma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Mouse/Rat CD31/PECAM‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3628) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CD31/PECAM‑1 at approximately 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|Detection of CD31/PECAM‑1 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Goat Anti-Mouse/Rat CD31/PECAM‑1 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF3628, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108).|
|Detection of CD31/PECAM‑1 in Rat Splenocytes by Flow Cytometry. Rat splenocytes were stained with Goat Anti-Mouse/Rat CD31/PECAM‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3628, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108).|
|CD31/PECAM‑1 in Mouse Embryo. CD31/PECAM‑1 was detected in immersion fixed frozen sections of mouse embryo (E13.5) using Goat Anti-Mouse/Rat CD31/PECAM‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3628) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to developing endothelium. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.|
PECAM-1 (Platelet-Endothelial Cell Adhesion Molecule-1), also known as CD31, is a 130 kDa type I transmembrane glycoprotein adhesion molecule in the immunoglobulin superfamily (1, 2). Expression is restricted to cells involved in circulation, especially endothelial cells, platelets, monocytes, neutrophils and lymphocyte subsets. PECAM-1 is concentrated at cell-cell junctions and is required for Transendothelial Migration (TEM) (1-3). The Extracellular Domain (ECD) of PECAM-1 has ten potential N-linked glycosylation sites and six C2-type Ig-like domains, the first of which is critical for adhesion and extravasation (3, 4). The cytoplasmic domain contains Immunoregulatory Tyrosine-based Inhibitory and Switch Motifs (ITIM, ITSM) that mediate both inhibition and activation via phosphotyrosine-mediated engagement of SH2-containing signaling molecules (1, 5). Metalloproteinase-mediated ectodomain shedding occurs during apoptosis (6) but increased serum PECAM-1 ectodomain in HIV and active multiple sclerosis occurs independent of apoptosis (7, 8). In humans, expression of six isoforms with exon deletions in the cytoplasmic domain is tissue- and stage-specific, but full-length PECAM-1 is predominant. A form lacking the ITSM predominates in mouse (9). Mouse PECAM-1 ECD shows 77%, 63%, 63%, 63%, and 61% amino acid (aa) identity with rat, human, canine, porcine, and bovine PECAM-1, respectively. PECAM-1 participates with other adhesion molecules in some functions, but is the critical molecule for TEM. Homotypic PECAM-1 adhesion in trans, combined with cycling of PECAM-1 to and from surface-connected endothelial cell vesicles, leads leukocytes across endothelial tight junctions (3, 10). Homotypic adhesion and signaling functions also strongly suppress mitochondria-dependent apoptosis (11). In platelets, PECAM-1 is necessary for limiting thrombus formation (12) and promoting integrin-mediated clot retraction and platelet spreading (13), but mechanisms for these phenomena are unclear. PECAM-/- mice are deficient in chemokine-mediated chemotaxis (14).
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Immunohistochemistry: Mouse/Rat CD31/PECAM-1 Antibody [AF3628]
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