advice

T. J. Murphy medtjm at bimcore.emory.edu
Fri Jul 21 14:23:50 EST 1995


In article savidge at unb.ca, savidge at unb.ca ("Dr. R. A. Savidge") writes:
>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!

oops!

>I don't have direct access to Swissprott or other data banks.  Can anyone
>confirm the homology interpretation?

yep, see gp:A03758 for an entry into the literature.

>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 don't suppose this piece of pine was used as an ice hockey stick before
it was sacrificed for science? 

>I am going to repeat the purification and, funds permitting, will attempt
>another sequence.  Does anyone have any other suggestions?

Imagine what the other guy is thinking: "How in the hell did I get a protein 
from a pine tree in my sample of purified human serum albumin??!!!???"

>**********************************************************************
>   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                          |




---


TJ Murphy
Asst. Professor	      
Dept of Pharmacology
Emory University School of Medicine   	    





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