In article <20030625122256.8089.qmail at ww02.hostica.com>, cvaravindhan at yahoo.co.uk wrote:
>>hi guys,
>i am working with a protein which is 10kda.i am not able see a band at the
> expected region in westernblotting-things which i hav used
>1.amersham ECL NCM -poresize 2u
>2.0.1%SDS in blot buffer
>3..63A for 30-45 min
>4.0.3% Tween-20 in PBS for wasing and blocking
> plz suggest me few things abt were i am going wrong.
As you don't mention controls, it is impossible to eliminate
any of the zillion possible reasons. However, my first reaction
is that the culprit is 0.1% SDS. 10 kDa is small enough to run
through membrane. On the other hand, at least something
should have remain bound, so you are also might have
problem with sensitivity.
DK
