Hi Tom,
in order to render your N-formyl-Met protein susceptible to sequencing, you
could try to incubate it with diluted acid (e.g. HCl), this should
hydrolyse the formyl group. The blot onto PVDF membrane should not give
rise to any problems, since PVDF is the membrane of choice for sequencing
of blotted proteins.
Depending on the sensitivity of your sequencer, a minimum amount of 10pmol
of blotted protein should be a reasonable bottom line.
Good luck and best regards,
Martin.
*****************************
Martin Kussmann, PhD
R&D, Protein Analysis /
Mass Spectrometry
CERBIOS-PHARMA S.A.
Via Pian Scairolo 6
CH - 6917 Barbengo (Lugano)
Switzerland
Phone: ++41 - 91 - 985 63 11
Fax: ++41 - 91 - 985 63 25
E-mail: rdbio2@cerbios.ch
-----Original Message-----
From: Tom Robinson [SMTP:trobinson@pdl.com]
Sent: lunedi, 3. aprile 2000 18:13
To: Recipients of ABRF List
Subject: N-formyl Met in N-terminal sequencing
Does anyone have any experience in rendering a protein with an N-terminal
N-formyl methionine residue amenable to sequencing? The protein is
blotted to PVDF.
Thanks in advance for any advice,
Tom Robinson
Protein Design Labs, Inc.
trobinson@pdl.com
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