Hi Mintu,
I suggest using purified whole protein for ESI-MS. You will get a series of
protein peaks (if the substitution is not 100%, which is often the case), each
differ for 47 Dalton. You can determine how many Se each peak corresponds to if
you know the molecular weight of your protein. Then you can integrate the peaks
to get an overall incorporation percentage.
Zhijie
From: Mintu Chandra
Sent: Monday, June 23, 2014 11:57 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] protocol for seleno-methionine incorporation...
Dear all,
Is there any standard protocol to know the number of incorporated
seleno-methionine in a protein ??? I am growing one of the protein in minimal
media for phase determination.
I am using mass spectrometry facility to know the incorporated
seleno-methionine but I wanted to know whether I should use the purified
protein or digested protein for mass spectrometry.
Thanks,
Mintu Chandra.
--
Mintu Chandra
Senior Research Fellow
IISER BHOPAL
mi...@iiserb.ac.in
+918085288853
