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 <[log in to unmask]> 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 <[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?? 

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WITH REGARDS
Rohit Kumar Singh
Lab. no. 430,
P.I. Dr. S. Gourinath,
School of Life Sciences,
Jawaharlal Nehru University
New Delhi -110067

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----------------------
Anirban Kundu

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WITH REGARDS
Rohit Kumar Singh
Lab. no. 430,
P.I. Dr. S. Gourinath,
School of Life Sciences,
Jawaharlal Nehru University
New Delhi -110067