thanks for the suggestion, is SDS will help in case of CAPS buffer? molecular weight of my protein is 40 kda. or can i use PO4 buffer for N-ter sequencing.
On Sat, Mar 21, 2015 at 12:42 PM, Anirban Kundu <anirbank...@gmail.com> wrote: > Try with 5 % methanol along with CHAPS buffer and the membrane should be > methanol charged > > On Fri, Mar 20, 2015 at 10:48 PM, rohit kumar <rohit...@gmail.com> wrote: > >> Dear all, >> >> sorry for off topic discussion. >> I am having trouble transferring protein onto PVDF membrane for >> N-terminal sequencing. When I used Tris-Glycine-Methanol in transfer buffer >> I got good blot, but when I used CAPS buffer (pH 11 set with 2N NaOH and >> 10% methanol) I did not get any blot. I tried transferring with variable >> current and durations (2hrs to overnight). Can anyone suggest what I might >> do?? >> >> -- >> WITH REGARDS >> Rohit Kumar Singh >> Lab. no. 430, >> P.I. Dr. S. Gourinath, >> School of Life Sciences, >> Jawaharlal Nehru University >> New Delhi -110067 >> > > > > -- > ---------------------- > Anirban Kundu > -- WITH REGARDS Rohit Kumar Singh Lab. no. 430, P.I. Dr. S. Gourinath, School of Life Sciences, Jawaharlal Nehru University New Delhi -110067
