2 bands very close in the expected place - (Mar/16/2016 )
Dear all,
I have done WB, but I did denaturing of my sample for only 3 minutes (I used sds-RIPA as lysis buffer).
So can these very close 2 bands be the native and denaturing protein, since I did not do the whole recommend time (5 minutes).
Thanks for any suggestion
Mady

is this a receptor or any membrane protein? have you ever done this and had only single band? Looking at the band pattern to me the two bands are a post-translational modified form of the same protein most likely Fully glycosyated vs non/partial glycosylated protein, or phosphorylation and non-phosphorylated. ofcourse you can confirm by denaturing at a longer time.
Thanks GNANA for ur comment, this is true a membrane protein.
anyway, I tried to increase the denaturation time as recommended, then I would update here .
Mady