Anti-CD31 antibody [MEC 7.46] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD31 antibody [MEC 7.46] - BSA and Azide free (AB264486)

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753)

ab7753 staining pan Cytokeratin in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min),permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab7753 at 1μg/ml and ab6046,Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117,Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080,Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS,Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD31 antibody [MEC 7.46] - BSA and Azide free (AB264486)

This data was developed using ab7388 the same antibody clone in a different buffer formulation.Flow cytometric analysis of / fixed / permeabilized B16-F0 (Mouse melanoma epithelial cell-like, Left) / bEnd.3 (Mouse brain endothelioma, Right) cells labelling CD31 with ab7388 at 1/1000 dilution (0.1µg) (Red) compared with a Rat monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab)2 Anti-Rat IgG Fc (Alexa Fluor® 488, ab150161) at 1/2000 dilution was used as the secondary antibody. Negative control : B16-F0.Gated on viable cells.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD31 antibody [MEC 7.46] - BSA and Azide free (AB264486)

This data was developed using ab7388 the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized bEnd.3 cells labelling CD31 with ab7388 at 1/50 (19.32 μg/ml) dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in bEnd.3 cell line. Negative control : B16-F0 (PMID : 8082649) ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.