Actin expression in protein extracted from tissue

Peter Ellis via methods%40net.bio.net (by pjie2 from cam.ac.uk)
Fri Apr 3 12:23:15 EST 2009


On Fri, 3 Apr 2009, Kamalian, Laleh wrote:
>
> Above all this method of extraction was 
> previously done by one of my colleagues and worked very well, it gave a 
> single band of b-actin as the normalizer.  I followed her instructions 
> exactly.  I tried mouse monoclonal anti GAPDH yesterday and it also gave 
> me nonspecific bands, although the actual band ( according to the size) 
> was much better than the actin's.   So is there any further thoughts?

Grab some tissue you have plenty of, soak it in RNALater and practice the 
extraction protocol until it works in your hands before moving on to your 
precious tumour samples.

Do be aware that RNALater-treated samples are only suitable for 
*denaturing* gels and applications.  The information leaflet is quite 
clear on this point.

Peter


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