Protein blotting to PVDF membranes

[David Martin x3175]

We routinely blot to PVDF but use 10mM CAPS pH 11 10% MeOH as the transfer
buffer in a mini protean cell at a current of 100 mA. It takes ca. 60-90
mins.

to stain, 30 secs in coomassie (1g/l 40% MeOH 10% HOAc) then destain in
40% MeOH 10% HOAc. repeat to enhance sensitivity (maybe use a 15 or 20 sec
stain)

we use ABI ProBlott.. and use the protocol in the instructions. Our peptide
chemist reckons if you can see the band, it'll sequence (unless it is 
blocked)

good luck

...david Martin


dmartin at uk.ac.crc