Dear Laura,

a few years ago - when I realised that mamalian cell expression media *without 
Met* costed much more than *with Met* because it is classified as custom media 
- I tried to persuade the rep that there was a market for it - to no avail.

I did then plan an experiment in which I would deplete the media of Met using 
this enzyme :


https://en.wikipedia.org/wiki/Methionine_gamma-lyase

and then supplement the Met-depleted media with Se-Met.

At the time I discussed purchase of Met gamma-lyase from:

Alpha Laboratories

Web: www.alphalabs.co.uk<http://www.alphalabs.co.uk/>

Before I could attempt the experiment - a Pt salt then came to my rescue and I 
was able to calculate initial phases by SAD with a diffraction experiment at a 
Pt-edge wavelength - so I cannot tell you if the idea of generating my own 
media would have worked.

Just in case it helps!

Ciao

Pietro


Pietro Roversi

Lecturer (Teaching and Research) https://le.ac.uk/natural-sciences/

LISCB Wellcome Trust ISSF Fellow

<https://bit.ly/2I4Wm5Z>https://le.ac.uk/liscb/research-groups/pietro-roversi


Leicester Institute of Structural and Chemical Biology
Department of Molecular and Cell Biology, University of Leicester
Henry Wellcome Building
Lancaster Road, Leicester, LE1 7HB
England, United Kingdom

Skype: roversipietro
Mobile phone  +44 (0) 7927952047
Tel. +44 (0)116 2297237



________________________________
From: CCP4 bulletin board <[email protected]> on behalf of LAURA DEL AMO 
MAESTRO <[email protected]>
Sent: 10 March 2020 17:40
To: [email protected] <[email protected]>
Subject: [ccp4bb] Expi 293 (Met-) expression medium for selenomethionine 
labeling


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<https://eur03.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.thermofisher.com%2Forder%2Fcatalog%2Fproduct%2FA4096701%23%2FA4096701&data=02%7C01%7Cpr159%40LEICESTER.AC.UK%7C15b8a2ea1a0045a5078c08d7c51ba404%7Caebecd6a31d44b0195ce8274afe853d9%7C0%7C1%7C637194594816063263&sdata=kFB5h5yHX8d%2FfCqjmrxkhwzsx6coT4S9odKupAvhELU%3D&reserved=0>.
 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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