Hello Bad news: Expi cells work poorly in non-Expi medium :( Good news: you can add SelenoMet and make partially seleniated protein, which is usually enough. Note that too much SeMet is pretty toxic, and that the toxicity is associated with BOTH enantiomers of the amino acid, so if you have the ability to use the L-SeMet pure as opposed to the more common L,D-mix then you can put in more. Either way, it works.
Expect your yield to suffer. Question: did you try iodine incorporation? You can do it on the level of pure protein, or crystals... Artem - Cosmic Cats approve of this message On Tue, Mar 10, 2020 at 1:51 PM LAURA DEL AMO MAESTRO <lam...@ibmb.csic.es> wrote: > Dear ccp4 community, > > I am in my last 3 months of my PhD, working at the Structural Biology Unit > at IBMB-CSIC in Barcelona. As my experimental phasing trials using various > heavy metals failed so far, we ordered two months ago Expi 293 (Met-) > expression medium for selenomethionine labeling (see the reference: > https://www.thermofisher.com/order/catalog/product/A4096701#/A4096701. > But we got now the notice that it will not be delivered until end of April. > > I had been working so hard on this project, and this medium is essential > to finished with the experimental part of my thesis. Thus, I was wondering > if someone in the community has a bottle (1L) of this expression medium > without methionine that I could borrow, and which I would return > immediately once our medium arrives. > > Also in case you have recommendations or other sources of a defined Se-Met > medium for Expi293 cells, please let me know. With kind regards from > Barcelona, and hope someone can help, > > > Laura > > --- > > Laura del Amo Maestro - Proteolysis Lab > SBU-Structural Biology Unit, IBMB CSIC > Barcelona, Spain, Europe > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 > ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
