2 replies to this topic

[FionaGould]

I am isolating Triton-insoluble and soluble proteins from mammalian cells, yet I cannot get the insoluble protein pellet to re-dissolve in anything, like TNE, TNE with Triton or the SDS-PAGE loading dye/buffer. This is a nightmare for loading onto the gel. Anyone got any ideas?

[milanoj]

Fiona,
Try SDS to 1% with increasing concentrations of DTT or some other reducing agent and heat to 70C. Are there many cys residues in your insolubles?

[sbidw]

Try placing your samples in SDS loading buffer, then in  a sonicating bath. It worked for a friend of mine.