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problem with Western blot loadig control


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#1 nixux

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Posted 19 March 2009 - 01:37 PM

Hi, everyone,

I just wanna ask a very basic question about the choice of loading control.

I want to detect an enzyme protein (estimate MW is around 30-45 KD) in bacteria with western blot, but I don't know what loading control could be used since according to my knowledge, actin, tubulin and GADDH is specific for eukaryotes.

I'll be very apprecitive of any suggestion.

Thanks a lot!

#2 Dr Teeth

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Posted 20 March 2009 - 07:32 AM

I don't know what is normally used, but FYI, bacteria possess homologues for both tubulin and actin (FtsZ and mreB, respectively).

See:
Prokaryotic origin of the actin cytoskeleton
FUSINITA VAN DEN ENT, LINDA A. AMOS & JAN LWE
Nature 413, 39-44 (6 September 2001)

Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley

#3 nixux

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Posted 23 March 2009 - 12:47 PM

Thanks a lot.

However, I guess it'll be better to use the loading control that was commonly used by others.

Has anyone ever used a loading control for the quantitative western blot detection of the bacteria protein?
I am waiting for any useful suggestions.




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