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." */
