Staining nitrocellulose

Curt Ashendel ashendel at
Tue Feb 28 08:55:35 EST 1995

On Mon, 27 Feb 1995 17:19:57 GMT, 
R. John Lye  <rjl6n at> wrote:

>> Same solution as for staining gels 1% dye in 30% MeOH 10% acetic acid 
>> (though the solvent concentrations can be varied somewhat).  Immerse 
>> nitrocellulose for 10 to 15 seconds with agitation then destain for 1 to 
>> 20 min in same solvent lacking dye, until background is acceptable. This 
>> may be done after blocking and probing, but probing does not work after 
>> dye staining. 
>This assumes that you're not blocking with a protein-containing sol'n
>(like BSA, serume, etc.), otherwise you end up with a uniformly
>blue blot.

This is not the case. I routinely stain my blocked (either milk or BSA) and 
probed blots and have had no problem seeing marker and sample bands. Yes, 
there is a higher level of background, but as long as a reasonable amount 
of protein was loaded, you can use this to check for equal loading and to 
locate markers.  The solvent slightly shrinks the NC, so exact positioning 
of the markers is usually not possible.

Curt Ashendel
Purdue University
West Lafayette, IN
ashendel at

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