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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 <[log in to unmask]> 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