Can I pre-stain Prot markers for W. Blot?

ijiwaru at wheel.dcn.davis.ca.us ijiwaru at wheel.dcn.davis.ca.us
Mon Jun 17 13:54:01 EST 1996


In article <4ppu7u$rt4 at panix.com>, iayork at panix.com (Ian A. York) wrote:

> In article <whoever-1306961429590001 at gaskins.animal.uiuc.edu>,
> Dave <whoever at wherever.here> wrote:
> >
> >I mistakenly purchased non-stained protein markers from Fisher for use in
> >Western blots.  Is there any way I can pre-stain them myself, and still
> >get good transfer to a nitrocellulose membrane?
> 
> First, if you want replies, you should probably change your e-mail address 
> to a valid one.  Second, in your position I wouldn't bother trying to 
> pre-stain your markers; transfer to unstained markers and then stain your 
> NC with india ink, exactly as described in Harlowe's antibody manual 
> (from Cold Spring Harbor Press, I hear a new version is coming out 
> soon).  The non-stained markers will almost certainly run more accurately 
> and tighter than do the prestained ones, and the time and effort in india 
> ink staining are trivial; you can also use much less marker.  
> 
> There's really very little advantage to using prestained marker in 
> Westerns over the india ink staining.
> 
> Ian
Ian,

no offense, but I have to disagree with your appraisal of pre-stained
markers.  I use then to gauge the length of my gel runs.  I have several
proteins that I have to deal with and they all run at various positions
and on different gel systems (I usually try to tailor my gel and run for
the protein I'm interested in).  For that reason I use pre-stained
markers.  I do have to say that the Bio-Rad kaleidascope markers look nice
all the way through to the transfer, but do seem to fade once they have
been on the membrane for a while.  I do agree that they run a little
fuzzier and not quite as "accurately" as unstained markers, but for what I
do, I think they are worth it.  YMMV

Lyle Najita
Plant Pathology
University of California - Davis



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