brush border membrane vesicle preparation

[Dr Engelbert Buxbaum]

teerachild at yahoo.com wrote:
> 
> Hi, I just wanna know something about brush border membrane vesicle preparation (BBMV). Can I purifiy BBMV by boiling of insect guts in SDS-PAGE sample buffer ? 

This would give a preparation which would also include proteins from a
lot of other compartments (gut proteins rather than brush border), all
of which would be denatured and non-functional. They would not be in
vesicles (closed membrane fragments) but in detergent micelles.

The usual procedure would be to homogenise the tissue (Potter-Elveijhem)
and to isolate the brush border membranes by a series of cenrifugation
steps. Use marker enzymes to characterise your fractions. 

Check books on basic biochemical techniques for further info, "Methods
in Enzymology" for specific methods.