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Prestained SDS-PAGE markers WITHOUT reducing agent

Prestained SDS-PAGE markers WITHOUT reducing agent - Protein Forum

Prestained SDS-PAGE markers WITHOUT reducing agent - Protein Forum


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  #1  
Old 03-17-2006, 04:21 PM
DA
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Default Prestained SDS-PAGE markers WITHOUT reducing agent



Dear all,
I am trying to find a company that sells prestained
SDS-PAGE markers without added reducing agent. All of
the vendors I have found included reducing agents in
their products.
Can someone point me in the right direction?
Thank you very much.
D. Andrews
University of Miami

--- Dr Engelbert Buxbaum
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  #2  
Old 03-17-2006, 07:43 PM
DK
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Default Prestained SDS-PAGE markers WITHOUT reducing agent

In article <[Only registered users see links. ].net >, DA <[Only registered users see links. ]> wrote:

Buy whatever prestained markers you like and run
them over little spin gel-filtration columns.
(Bunch of companies sell them and they are trivial to
make out of blue tip, for example). Volume change
is minimal and the protein recovery is always >80%.

Sephadex G-25 is most common but I like
Bio-Rad's P6 better.

DK
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Old 03-18-2006, 06:55 PM
Allison
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Default Prestained SDS-PAGE markers WITHOUT reducing agent

If you leave a lane free between the markers and your non-reduced
samples you don't need to remove the reducing agent. I have done this
many times.
Allison

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  #4  
Old 03-19-2006, 05:50 PM
Amanda Gillespie
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Default [Protein-analysis] Re: Prestained SDS-PAGE markers WITHOUTreducing agent

Yes, this is true, but sometimes, depending on the composition of the
markers, the non-reduced forms can run differently to the reduced forms, so
for accurate MW measurement you need to be sure that there is no difference
before resorting to this method - at least, that's been my experience...

Amanda


On 3/18/06, Allison <[Only registered users see links. ]> wrote:
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  #5  
Old 03-21-2006, 08:45 PM
Amanda Gillespie
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Default [Protein-analysis] Re: Prestained SDS-PAGE markers WITHOUTreducing agent

I use BioRad's kaleidoscope markers on my non-reduced zymograms and reverse
zymograms all the time (testing for gelatinase activity in my samples) and
they have never given me any problems - I don't even have to leave a lane
between the marker and the sample - they don't seem to leak any kind of
interfering reducing agent.

I know this probably doesn't answer your question, but if you're looking for
markers to use with non-reduced samples, that's what I use without any
problems...

Amanda Gillespie
Postgraduate student - Biochemistry and Immunology
UKZN
South Africa

On 3/20/06, DA <[Only registered users see links. ]> wrote:



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