The role of DTT in cytosol extraction - (Oct/08/2005 )

Hallo everyone!
I am just presenting my research study for the MSc degree on Monday and I have a small question: what is the role of dithiothreitol in cytosol extraction? Is it only for reduction of molecules? Thanxx :

-paixtaki-

DTT is used to reduce proteins. Namely disulfide bonds.

-pBluescript-

DTT and b-mercapto are also some of the more "fragrant" things you get to work with

-Rafflez-

Nothing like the smell of BME in the morning.

-pBluescript-

hem... TEMED is not bad too

-fred_33-

DTT is used to reduce disulfide bonds from proteins that are already denatured preventing them to react with others and form aggregates.
It also helps maintain nuclear membrane integrity during cell fractionation, but not sure how.

-dapo-

its easier to use DTT compared to B-mercaptoetoh since the latter actually has a buffering effect, plus i believe that its more MALDI compatible with DTT.

-mices-

QUOTE (dapo @ Oct 14 2005, 09:22 AM)

DTT is used to reduce disulfide bonds from proteins that are already denatured preventing them to react with others and form aggregates.
It also helps maintain nuclear membrane integrity during cell fractionation, but not sure how.

Thanks for this tip: I'm actually using a ChIP protocol in which I have to incubate cells in 100mM Tris-HCl [pH 9.4] plus 100mM DTT before collecting nuclei... and I was wondering why...

-HP1-

which is a more effective reducing agent DTT, Tri Butyl phosphine b-Mercaptoethanol, Tris (2-Carboxyethyl)- Phosphine hydrochloride with respect to 2 D Gel Electrophoresis and MALDI

-SHIVA KESHAVA-