[diablo]

Hi,

is anyone of you familiar with Peptidesynthesi/FMOC/ BOC strategies, if yes please help me ^^

i got ("dried" in speedvac) which are protected with BOC & FMOC and i wanted to kick the FMOC --> i added 20%Piperidine (H2O) and the FMOC's were kicked.

Now I got the same peptides (diluted in DMSO) and i added 20%Piperidine (now with DMSO --> not H20) and my resluts (MS/MS)  are telling me that it doesn't work...  now i have to find the failure:

1) the peptide concentration was very low..
2) DMSO/Piperidine doesn't work ? ( i don't think so)
3) or does the BOC somehow interfere with Piperidine ? ( I BOCylated the peptide myself (BOC in DMSO, Peptide in DMSO)... after incubation for 1 hour i added the Piperidine/DMSO solution to the same sample...

Edit: i didn't prepared the Piperdine/DMSO solution some days ago ( not fresh...) does this matter?

thx & Greets
Diablooo

[Arkcon]

Well, here's a link {click} to a number of FMOC deblocking procedures that you can try.  Basically, DMSO and piperidine are pretty stable in air, but I have seen preps fail with very old solutions, a few days might not be too old, but maybe you should start fresh and see.