protocol for methionine incorporation

Nick Theodorakis nick_theodorakis at hotmail.com
Sun Jan 11 17:23:27 EST 2004


On 11 Jan 2004 15:16:36 -0000, viegas at embl.de wrote:

>
>Hi!
>I wonder if someone has a protocol for [35S]methionine incorporation to be used 
>in Hela cells. In particular, I am interested in the amount of uCi to use per 
>ml. and per number of cells. Secondly, I´d like to now if the cells have to be 
>in confluence for this essay or not. Thank you very much in advance,
>                                        Marcelo Viegas
>

I've used as few as 100-200 cells, or up to 10 million. And from 5 uCi
to over 200 uCi of [35S]methioine. It depends on what you are trying
to do. Are you looking at general labelling of proteins, or trying to
immunoppt. a specific (perhaps rare) protein?

Nick

-- 
Nick Theodorakis
nick_theodorakis at hotmail.com
nicholas_theodorakis [at] urmc [dot] rochester [dot] edu 



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