EEA1 antibody (rabbit 243)-produced by Ian Mills,1998. obtain antibody from MC.

protocols used successfully by IM.

Western Blotting

1. Block with PBS/5%Marvel for 30min.

2. Primary (anti-EEA1 antibody) to be used at 1:1000 in PBS/5% Marvel. One hour incubation at room temp.

3. Five washes (5min each) with PBS/5% Marvel/0.05% Tween.

4. Secondary anti-rabbit HRP (in fridge) to be used at 1:1000 in PBS/5% Marvel. One hour incubation.

5. Five washes (5min each) with PBS/5% Marvel/0.05% Tween. Then wash with PBS.

6. ECL development