Dear Artem,

By work poorly do you mean, not as well and are you referring to all Expi cells?

We have adapted ExpiCHO cells to another medium and although we see drastic 
decrease from using the complete Expi system (x15 volumetric yield), it was 
still considerably higher than our previous cells in the same medium (x5 
volumetric yields).

Will soon try with ExpiSf9 and baculovirus to see if they also perform 
similarly.

Best,

Kelvin

On Mar 11, 2020, at 5:49 PM, Artem Evdokimov 
<artem.evdoki...@gmail.com<mailto:artem.evdoki...@gmail.com>> wrote:

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

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On Tue, Mar 10, 2020 at 1:51 PM LAURA DEL AMO MAESTRO 
<lam...@ibmb.csic.es<mailto: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

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