Help in Western blot

John K. Troyer jtroyer at
Mon Sep 18 11:57:07 EST 1995

On 15 Sep 1995, Giorgio Spagnol wrote:

> Dear fellow researchers,
> I have low yeld in transfering a glycoprotein to nitrocellulose membranes
> for immunoblotting. It is 100 kD and 30% carbohydrate by weight, and
> quite acidic (PI around 4.4). I would be grateful for any suggestion.
> Cordially
> Giorgio.


Methanol in the transblot buffer can retard the transfer of high 
molecular weight (>100 kd) species and particularly, glycoproteins.  
Reduction or total elimination of the methanol may greatly improve your 
transfers.   Hope this helps.



John K. Troyer, PhD
University of Maryland School of Medicine
Department of Biological Chemistry
(401) 706-7518
jtroyer at

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