CD31/PECAM-1 Antibody (3155B) [Unconjugated]

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.

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.

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.

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.

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.

• Reactivity: Mu
• Applications: IHC, mIF
• Clone: 3155B
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

CD31/PECAM-1 Antibody (3155B) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line, NS0-derived mouse CD31/PECAM-1
  Glu18-Lys590
  Accession # Q08481
• Specificity: Detects recombinant mouse CD31 in Direct ELISA.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 3-25 ug/mL
  • Multiplex Immunofluorescence 10 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Reconstitution Instructions: Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CD31/PECAM-1 Antibody (3155B) [Unconjugated]

• adhesion molecule
• CD31 antigen
• CD31
• CD31/EndoCAM
• EndoCAM
• FLJ34100
• FLJ58394
• GPIIA'
• PECA1
• PECAM1
• PECAM-1
• PECAM-1, CD31/EndoCAM
• platelet endothelial cell adhesion molecule
• platelet endothelial cell adhesion molecule-1
• platelet/endothelial cell adhesion molecule

Background

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).

1. Ilan, N. and J.A. Madri (2003) Curr. Opin. Cell Biol. 15:515.
2. Xie, Y. and Muller, W.A. (1993) Proc. Natl. Acad. Sci. USA 90:5569.
3. Liao, F. et al. (1997) J. Exp. Med. 185:1349.
4. Nakada, M.T. et al. (2000) J. Immunol. 164:452.
5. Chemnitz, J.M. et al. (2004) J. Immunol. 173:945.
6. Ilan, N. et al. (2001) FASEB J. 15:362.
7. Eugenin, E.A. et al. (2006) J. Leukoc. Biol. 79:444.
8. Losy, J. et al. (1999) J. Neuroimmunol. 99:169.
9. Wang, Y. et al. (2003) Am. J. Physiol. Heart Circ. Physiol. 284:H1008.
10. Mamdouh, Z. et al. (2003) Nature 421:748.
11. Gao, C. et al. (2003) Blood 102:169.
12. Falati, S. et al. (2006) Blood 107:535.
13. Wee, J.L. and D.E. Jackson (2005) Blood 106:3816.
14. Wu, Y. et al. (2005) J. Immunol. 175:3484.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.