SDS is preferred for fast transfer. On Sat, Mar 21, 2015 at 4:24 PM, rohit kumar <rohit...@gmail.com> wrote:
> 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 > -- ---------------------- Anirban Kundu
