Hello Ship,
I do not have your system but the following should help:
run your nanospray in 5% formic acid (FA), 50% methanol in water. How to
get your sample in this buffer? A few possibilities: 1st: add FA to your
digest, dry in speedvac, solubilize in buffer and run your NanoMS
experiment if it is still not working try: 2nd: use ZipTip from Millipore
(C18mycro is best for high sensitifity) with there protocol or 3rd: you can
also use Poros P2 in a gelloading tip as your home made RP-column to desalt
your peptides.
Greetings Daniel
>Hello,everybody, we need some help. We have been trying to run some tryptic
>digests on our Bruker Esquire LC-MS system using their nanospray source. We
>can
>get reasonably good results on pure peptides but with tryptic digests we are
>not getting good results at all. We do our digests in 2mM NH4HCO3 (pH
>7.5-8.0). Is our
>failure due to the presence of buffer (NH4HCO3) in the digest? If so or
>even otherwise does anybody have a suggestion regarding this problem. We do
>not have too
>much experience in this area and we would welcome any help that we can get.
>Thanks.
>
>
>
>Shib
>
>
>
> Shib Prasad Dutta, Ph.D.
> Roswell Park Cancer Inst.
> Elm & Carlton Sts.
> Buffalo, New York 14263.
> Shib.Dutta@roswellpark.org
Dr. Daniel Hess
Friedrich Miescher Institut
Maulbeerstr. 66
CH-4058 Basel
Switzerland
Phone: Office ++41 61 697 85 44, Lab ++41 61 697 68 68
FAX: ++41 61 697 39 76 e-Mail: dhess@fmi.ch
This archive was generated by hypermail 2b29 : Fri Aug 25 2000 - 08:35:04 EDT