[Drosophila] Protein Extract from Larvae

Kathleen Gajewski via dros%40net.bio.net (by sa08383 from wotan.mdacc.tmc.edu)
Tue Mar 18 12:04:10 EST 2008


Ivy, Have you tried freeze-drying?  I've done Westerns, 2D gels and RNA 
preps from freeze tried tissues with good results.  Here's the 
reference:

Fujita S.C., Inoue H., Yoshioka T., Hotta Y., 1987.  Quantitative 
tissue isolation from Drosophila freeze-dried in acetone.  Biochem. J. 
243, 97-104.


Good luck,

KG


On Feb 25, 2008, at 12:11 AM, Ivy McDaniel wrote:

> Hello Drosophila Community,
>
> I'm looking for a good protocol, or even just general tips for making 
> protein extracts from third-instar larvae and adult flies. I'm dealing 
> with a very large protein, and I'm having a lot of trouble avoiding 
> degradation. Grinding adults/larvae/salivary glands directly in 
> boiling SDS loading buffer hasn't worked, nor has one protocol where I 
> included protease inhibitors at every step. Any advice at all would be 
> greatly appreciated!
>
> Thanks,
> Ivy
>
>
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