multimer on denatured SDS-PAGE?

Cornelius Krasel krasel at wpxx02.toxi.uni-wuerzburg.de
Sun May 3 11:41:42 EST 1998


Rick00100 <rick00100 at aol.com> wrote:
> I'm a newbie in protein work and Western, and recently I'm trying to raise
> antibodies to my protein, which is about 22kd.  I got some polyclonal Ab,
> and when I do a Western with CHO cells-transfected protein, I would not see
> a band with one of my Ab.  However, after deglycosylating the cell lysates,
> I can see a band at around 40kd, and a faint band at 22kd.

It's possible that the 40 kDa band is a dimer. Since your protein is
a membrane protein (why else would you want to deglycosylate it), this
is not entirely surprising. Sometimes it helps to omit the boiling step
(at least this is true for some G protein-coupled receptors).

--Cornelius.

-- 
/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
/* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP4 */
/* "Science is the game we play with God to find out what His rules are."  */



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