Mouse CD31/PECAM-1 Antibody Summary
Glu18-Lys590
Accession # Q08481
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of CD31 in Frozen Mouse Brain Cortex via seqIF™ staining on COMET™ CD31 was detected in perfusion fixed-frozen sections of mouse Brain Cortex using Rabbit Anti-Mouse CD31, Monoclonal Antibody (Catalog #MAB11720) at 10ug/mL at 27° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent preprocessing by incubating tissue with Multi Staining Buffer (Lunaphore Catalog # BU06) for 5minutes at room temperature followed by a 20 minute incubation in Tris-Buffered Saline + 0.2% Triton at room temperature. Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm of endothelial cells. Protocol available in COMET™ Panel Builder.
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Detection of CD31 in Frozen Mouse Hippocampus via seqIF™ staining on COMET™ CD31 was detected in perfusion fixed-frozen sections of mouse Hippocampus using Rabbit Anti-Mouse CD31, Monoclonal Antibody (Catalog #MAB11720) at 10ug/mL at 27° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent preprocessing by incubating tissue with Multi Staining Buffer (Lunaphore Catalog # BU06) for 5minutes at room temperature followed by a 20 minute incubation in Tris-Buffered Saline + 0.2% Triton at room temperature. Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm of endothelial cells. Protocol available in COMET™ Panel Builder.
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Detection of CD31 in Frozen Mouse Brain Medulla via seqIF™ staining on COMET™ CD31 was detected in perfusion fixed-frozen sections of mouse Brain Medulla using Rabbit Anti-Mouse CD31, Monoclonal Antibody (Catalog #MAB11720) at 10ug/mL at 27° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent preprocessing by incubating tissue with Multi Staining Buffer (Lunaphore Catalog # BU06) for 5minutes at room temperature followed by a 20 minute incubation in Tris-Buffered Saline + 0.2% Triton at room temperature. Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm of endothelial cells. Protocol available in COMET™ Panel Builder.
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Detection of CD31/PECAM‑1 in Mouse Kidney. CD31/PECAM‑1 was detected in perfusion fixed paraffin-embedded sections of mouse kidney using Rabbit Anti-Mouse CD31/PECAM‑1 Monoclonal Antibody (Catalog # MAB11720) at 5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane of endothelial cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
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Detection of CD31/PECAM‑1 in Mouse Thymus. CD31/PECAM‑1 was detected in perfusion fixed paraffin-embedded sections of mouse thymus using Rabbit Anti-Mouse CD31/PECAM‑1 Monoclonal Antibody (Catalog # MAB11720) at 5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane of endothelial cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: CD31/PECAM-1
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. CD31 is concentrated at cell-cell junctions and is required for transendothelial migration (TEM) (1‑3). The extracellular domain (ECD) of CD31 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 CD31 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 CD31 is predominant. A form lacking the ITSM predominates in mouse (9). Mouse CD31 ECD shows 77%, 63%, 63%, 63% and 61% amino acid (aa) identity with rat, human, canine, porcine and bovine CD31, respectively. CD31 participates with other adhesion molecules in some functions, but is the critical molecule for TEM. Homotypic CD31 adhesion in trans, combined with cycling of CD31 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, CD31 is necessary for limiting thrombus formation (12) and promoting integrin-mediated clot retraction and platelet spreading (13), but mechanisms for these phenomena are unclear. CD31-/- mice are deficient in chemokine-mediated chemotaxis (14).
- Ilan, N. and J.A. Madri (2003) Curr. Opin. Cell Biol. 15:515.
- Xie, Y. and Muller, W.A. (1993) Proc. Natl. Acad. Sci. USA 90:5569.
- Liao, F. et al. (1997) J. Exp. Med. 185:1349.
- Nakada, M.T. et al. (2000) J. Immunol. 164:452.
- Chemnitz, J.M. et al. (2004) J. Immunol. 173:945.
- Ilan, N. et al. (2001) FASEB J. 15:362.
- Eugenin, E.A. et al. (2006) J. Leukoc. Biol. 79:444.
- Losy, J. et al. (1999) J. Neuroimmunol. 99:169.
- Wang, Y. et al. (2003) Am. J. Physiol. Heart Circ. Physiol. 284:H1008.
- Mamdouh, Z. et al. (2003) Nature 421:748.
- Gao, C. et al. (2003) Blood 102:169.
- Falati, S. et al. (2006) Blood 107:535.
- Wee, J.L. and D.E. Jackson (2005) Blood 106:3816.
- Wu, Y. et al. (2005) J. Immunol. 175:3484.
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