I am embarrassed to communicate what follows, but I am seeking advice with
the thought that an interpretation mistake has been made somewhere that can
be explained by experienced protein researchers.
A postdoc of mine isolated a protein from a pine tree by a series of
fractionation steps including preparative IEF, SDS-PAGE and electroblotting
to PVDF membrane, a band from the latter being N-terminal sequenced. A
signal corresponding to about 5 pmoles was obtained. The sequence was
NH2-DAHKSEVAHRFKDLGEEN and, according to the sequencer, this has 100%
homology to human serum albumin as provided by the Swissprott data bank!
I don't have direct access to Swissprott or other data banks. Can anyone
confirm the homology interpretation?
We are certain that we did not use any type of serum albumin at any stage
of protein purification. Sterile gloves were worn for the SDS and
electroblotting procedures. The sequencer suggested that the gel may have
been contaminated by blood, but that there would have had to have been a
lot to obtain such a nice signal. We are certain that there were no cuts
on fingers and that blood cannot be the explanation.
I am going to repeat the purification and, funds permitting, will attempt
another sequence. Does anyone have any other suggestions?
**********************************************************************
Rod Savidge, PhD | E-mail: savidge at unb.ca
Faculty of Forestry and \|/
Environmental Management \ | / Phone: (506) 453-4919
University of New Brunswick _\/ | \/_
Fredericton, NB CANADA \|/ Fax: (506) 453-3538
E3B 6C2 |
