what would happen if i dilute protease inhibitor too much in cell lysate? - (Apr/01/2008 )

my protease inhibitor stock solution is 100x concentrated. but when i prepared the lysis buffer for cell lysate, i diluted it 1000 times and didn't noticed until i've finished all the extraction procedures! - - annoyed by myself.

so do i need to redo the extraction? or i could do a protein assay to get the concentration and see if the proteins are degraded?

thanks~

the protein assay won't tell you the condition of your protein extract. you will need to see them on a gel and compare them to an extract that was done correctly.

however, the inhibitors are usually in excess of the functional concentration and you may be lucky enough that your extract is okay (but add more inhibitor to adjust to the proper level, after you find out that it is okay (so you don't waste them)).

Yes, but what does the gel look like? Does your protein look the right size? If not, then re-do the prep. But if it looks as though your protein is safe, finish the prep and try a few expts. Then go to the bathroom, look yourself in the mirror, call yourself a thousand silly names, and make yourself promise to never do it again.