Invitrogen

CD31 Monoclonal Antibody (HEC7)

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated. View Details

14 Published Figures

28 References

View all (161) CD31 (PECAM-1) antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

CD31 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of CD31 (green) showing staining in the membrane and cytoplasm of THP-1 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CD31 monoclonal antibody (Product # MA3100) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

CD31 Antibody in Immunocytochemistry (ICC) — Immunofluorescence analysis of Platelet endothelial cell adhesion molecule was performed using 70% confluent log phase HUVEC cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CD31 Monoclonal Antibody (HEC7) (Product # MA3100) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing Cell Junctional localization. Panel e represents no expression in MDA-MB-231. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

CD31 Antibody in Western Blot (WB) — Western blot was performed using Anti-CD31 Monoclonal Antibody (HEC7) (Product # MA3100) and a 130kDa band corresponding to Platelet endothelial cell adhesion molecule was observed across cell lines tested. Membrane enriched extracts (40 µg lysate) of HUVEC (Lane 1), HEL 92.1.7 (Lane 2) and MDA-MB-231 (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).

CD31 Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of CD31 in PBMC cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a CD31 monoclonal antibody (Product # MA3100) at a dilution of 0.5 µg/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

CD31 Antibody in Relative expression — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell lines tested owing to their inherent genetic constitution. Relative expression of Platelet endothelial cell adhesion molecule was observed to be high in HUVEC in comparison to no expression in MDA-MB-231 using Anti-CD31 Monoclonal Antibody (HEC7) (Product # MA3100) in Western Blot (Ref :https://www.nature.com/articles/onc2013214). {RE}

CD31 Antibody — Fig. 2 TNBC cells induce PSMA expression on endothelial tubes. Fluorescence microscopy analysis of tube-associated expression of CD31 and PSMA on HUVEC cells incubated in a EBM-2 growth media supplemented with endothelial growth factors, b MDA-MB231-conditioned medium, and c MCF-7-conditioned medium

Product Details

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

3-5 µg/1x10^6 cells

View 2 publications 2 publications

Immunocytochemistry (ICC)

1:50-1:500

View 3 publications 3 publications

Immunofluorescence (IF)

1:50-1:500

View 10 publications 10 publications

Western Blot (WB)

1:1,000

View 2 publications 2 publications

Immunohistochemistry (IHC)

View 4 publications 4 publications

Neutralization (Neu)

View 5 publications 5 publications

ELISA (ELISA)

View 1 publication 1 publication

Immunoprecipitation (IP)

View 1 publication 1 publication

Product Specifications

Species

Bovine, Human

Published species

Artificial Control, Bovine, Human, Mouse, Rat

Expression System

Mouse / IgG2a

Class

Monoclonal

Type

Antibody

Clone

HEC7

Immunogen

Human PECAM-1

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein G

Storage buffer

PBS

Contains

no preservative

Storage conditions

-20°C

RRID

AB_223516

Product Specific Information

MA3100 targets CD31 in ICC/IF and ACS applications and shows reactivity with Human and Bovine samples.

The MA3100 immunogen is human PECAM-1.

MA3100 detects CD31 which has a predicted molecular weight of approximately 80 kDa.

This product has been tested for endotoxins by limulus amoebocyte lysate (LAL) assay and contains an endotoxin concentration of less than or equal to 10 endotoxin units per milligram (EU/mg).

Target Information

CD31 (platelet endothelial cell adhesion molecule-1, PECAM-1) is an inhibitory coreceptor involved in regulation of T cell and B cell signaling by a dual immunoreceptor tyrosine-based inhibitory motif (ITIM) that upon associated kinases-mediated phosphorylation provide docking sites for protein-tyrosine phosphatases. CD31 is expressed ubiquitously within the vascular compartment and is located mainly at junctions between adjacent cells. N-terminal Ig-like domain of CD31 is responsible for its homophilic binding, which plays an important role in cell-cell interactions. CD31 is a multifunctional molecule with diverse roles in modulation of integrin-mediated cell adhesion, transendothelial migration, angiogenesis, apoptosis, negative regulation of immunoreceptor signaling, autoimmunity, macrophage phagocytosis, IgE-mediated anaphylaxis and thrombosis. It is one of key regulatory molecules in vascular system.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: CD31; CD31 antigen; EndoCAM; GPIIA'; PECA1; PECAM-1; Platelet endothelial cell adhesion molecule; platelet/endothelial cell adhesion molecule (CD31 antigen); platelet/endothelial cell adhesion molecule 1

Gene Aliases: CD31; CD31/EndoCAM; endoCAM; GPIIA'; PECA1; PECAM-1; PECAM1

UniProt ID: (Human) P16284

Entrez Gene ID: (Bovine) 282303, (Human) 5175

Molecular Function: cell adhesion molecule