Nuclear extract without DTT? - (Mar/20/2007 )

Dear all,
I have a very basic question concerning nuclear extracts. I made prepared nuclei from insect cells but missed
to add 1 mM DTT to the lysis buffer. Does this somehow affect protein stabiity (the nuclei are in the -80 freezer)?

Cheers,

speedy

---

well DTT is a reducing agent. So adding it when your samples are thawed should be good (at least not damaging)

---

DTT works as anti-oxidant as pO2 increases during lysis and the intracellular anti-oxidant system has broken down; so you may have some more oxidized (= damaged) molecules in your preparation as if you have used DTT; as fred_33 said, during freezing it is not so important

---

Thank you, guys,
you helped me a lot!

---