Transfer of large proteins (>500 kD) to nitrocellulose. How?

Bob Obar 102063.2640 at CompuServe.COM
Sun Dec 29 19:14:24 EST 1996


Gunther -
  I would agree with Anton's recommendations, but if I were you I 
would ABSOLUTELY avoid the methanol (MeOH) in the transfer buffer!! 
It will almost definitely screw up the transfer of any large (i.e. 
>30kDa) polypeptide).  When I worked with microtubule-associated 
proteins such as MAP1A, MAP1B, MAP2, dynein heavy chain (all between
200kDa and 500kDa) we found that methanol always hurt the transfer 
efficiency.  You might also want to look up what the mucin people 
do, since those proteins are huge and very poorly behaved.  Best of 
luck (and you needsome  luck).

Bob Obar
102063.2640 at compuserve.com



More information about the Methods mailing list