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2D gelelectrophoresis: prestaining of proteins - (Nov/26/2008 )

to find differences in differently treated cell, f.i., total protein of lysates can be stained before separation of proteins... does anyone know which chromophores are suitable? I like to label an enzyme inhibitor with such a chromophore...

-The Bearer-

I dint understand, are you talking about DIGE?

-amj2-

prestaining is not a good idea for ief. pI may be altered by the stain.

prestaining will also increase the size of the protein and will affect mobility in the second dimension.

it is best to run the 2d then stain.

-mdfenko-

QUOTE (amj2 @ Dec 11 2008, 08:40 AM)
I dint understand, are you talking about DIGE?


yes, you ┬┤re right, but I forgot about it; now I remember I had 10 years ago or so a demo cd for the sofware

pre-staining with Cy-2, Cy-3 or Cy-5

-The Bearer-

yes you can stain your proteins with cy dyes but i thought that was to stain the lysates and then look at the differentially expressed proteins in fluorescence......i don't know whether your enzyme would need some specific dye, have you tried with any of these cy dyes?

-amj2-