At 10:15 16.08.00 -0600, you wrote:
> Hi to chop off the proteins. Peptide fragments will be applied to
>MALDI-TOF followed by digestion. Here some candidates: CNBr
>Endoproteinase Arg C Endoproteinase Asp N Endoproteinase Lys C V8
>(D,E) V8 (E) All above are available options from "ProFound" search
>Engine. (http://prowl.rockefeller.edu/cgi-bin/ProFound) Which company could
>you recommend to purchase it, also? Thanks in advance, KH Kim
>Chemical Engineering Dep. Colorado State University
>khkim@lamar.colostate.edu 970-491-1192
Hi,
I would recommend either Lys-C or V8 in bicarbonate (E-specificity). Both
work well within the gel. V8 is much cheaper than Lys-C and has different
cleavage sites than trypsin, which makes it (from my point of view) more
favourable than Lys-C. CNBr has the disadvantage of generating relatively
few and large fragments. Sometimes it can be an advantage if the protein of
interest is resistant to proteolytic cleavage, but most of the time, the
more fragments you get, the better for the search. Another problem with
CNBr ist the formation of different products. The cleavage will almost
always yield a mixture os homoserine and homoserine lactone, which will
complicate the analysis.
Hope this helps,
Marcus
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Dr. Marcus Macht
Universit”t K–ln
Zentrum f¸r molekulare Medizin
Zentrales Servicelabor
Joseph-Stelzmann-Str. 52
50931 Koeln
Tel.: +49 221 478-6995
Fax: +49 221 478-6977
e-mail: Marcus.Macht@uni-koeln.de
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