DTT in lämli buffer - (Dec/11/2006 )

Hi,


I use DTT in my lämli buffer for denaturating protein. Once I forgott to previous mix my lämli buffer with DTT bevor to add it to the cell lysat. So what I did was to mix lämli (without DTT) to my cell lysat to cook it and the I added some DTT and loaded on the SDS PAGE. I obseved a change in the consistance of my cell lysat. Could somebody explain me why?

1) should I have boil the sample again after adding DTT? and if yes why?
2) Is the DTT concentration important and what happen when I have too less DTT or too much DTT?

Thanks

cuca

I would have boiled after DTT, but I can't explain you why you observe this difference

Good to know !