EEA1, Abcam, ab2900

This antibody works very well for immunofluorescence staining in mouse (b.End3) and human (hCMEC/D3) brain endothelial cell line. Dilution 1:300. Fixation was done with 4% paraformaldehyde and cells were permeabilized with 0.5% Triton X-100.

Producer: Abcam
Catalog No: ab2900
Used dilution: 1:1000 (WB)
Ab name: EEA1
Secondary Ab: Anti-Rabbit HRP, Cell signaling 7074 (1:2000)
Cell line: primary porcine brain endothelial cells, bEND3
Host organism: Rabbit
Character: primary, Polyclonal

Antibody tested with primary porcine and bEND3 cell lysis.
In both cases the ab produced a single strong band at the appropriate size.

Producer: Abcam
Catalog No: ab2900
Used dilution: 1:200; 1:500; 1:1000 (IF)
Cell line: primary porcine brain endothelial cells, primary bovine brain endothelial cells
Host organism: Rabbit
Character: primary, polyclonal

Antibody tested in primary porcine and bovine brain endothelial cells. In both cases working fine giving clear dot/speckle-like signal. At higher dilutions signal significantly weaker. For IF as a secondary antibody Invitrogen goat anti-mouse Alexa Fluor 568 (1:350 or 1:200) was used. Cells fixed with 4% PFA, permeabilized with 0.1% Triton-X100, blocked with 2% BSA.

Not good for murine cell lines. Only very weak staining. CAnnot be recommended.

We used it in Western blotting on sucrose gradient fractions.

It recognizes a very faint band of the correct size at the expected density. A stronger (but still weak) double band is observed in the post nuclear supernatant of 293 cells.

Dilution 1:1000