protein isolation

Model Organism: Worms Methods & Reagents
1

Hi,

Can someone tell me about protein isolation from worms? How many worms do you typically use to get sufficient amount for downstream applications such as western? Also, what concentration of extracts do you typically get, someone who does this in cell culture system said they get at least 6-8ug/ul from a million cells. What volume of worms should I use to get about this concentration?

Thanks

2

Hi,

this was discussed recently and I put some figures here;

http://forums.wormbase.org/index.php?topic=1750.msg3975#msg3975

You can work out the necessary details for your Westerns from there.

Steve

3

You might want to look at this one too:

http://forums.wormbase.org/index.php?topic=1682.msg3771#msg3771

4

hi joan,
for westerns, i just transfer 5 worms to a standard SDS lysis-buffer, sonicate, boil and load the whole stuff onto a SDS-gel. that’s enough for whole cell extract immunoblot analysis.
cheers, oldcoffeee