2 replies to this topic

[AnnaSh]

Hi,
I've a question, that could result trivial to lot of you. I have to treat cells plated in small dishes (dia 3.5 cm) for very short time periods- 1-2 min- prior to lyse them for western blot assays. Can I stop treatments simply by transferring dishes on ice and immediately removing culture medium? Is it correct?
Thanks in advance for your replies

[bob1]

You can do it that way, you might want to wash them in ice-cold PBS first just to remove any residual drug and to instantly lower the temperature, remove this and then put in cold lysis buffer.  Be aware that this sort of thing also activates a whole heap of stress responsive genes, especially those containing cold-shock domains.

[AnnaSh]

Thanks for your reply! I think I'll do that way, also considering that transcriptional responses require more than a few minutes to occur.
Bye!