protein tranfer from gel to membrane, reversible ? - (Apr/03/2006 )

hi

while doing western transfer , i realized that i had kept the membrane at the cathode and gel near the anode ,, after half hour transfer i reversed and again trasfered
what are the chances that protein will stick to the membrane??

used nitrocellulose membrane
wet transfer , 350mA

do u think i should continue or start over again??
thanks

hi phytoviridae ! mistakes do happen . stain your membrane with ponceau S stain and check the amount tranfered. you might have definitely lost some of your protein, but there are fair chances that some protein might have been transfered on to the membrane. you didnt mention the type of apparatus you used. traner rate is high in semi dry apparatus and less time is needed for the transfer of proteins.......... that means in your case it is more protein loss.
any way all the best

hi
thank you for replying

i am not doing semi dry transfer but the immersion type
and there is no ponceau stain right now in lab ,,i wil have to borrow it from somewhere

anyways i will repeat it again ,, for which i will have to make some adjustments with labmates since there is only one appratus and many people need to use it


thanks again