Help in Western blot

John K. Troyer jtroyer at umabnet.ab.umd.edu
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.

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.

Regards,

John


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



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