MANUALPAGE 1

EXPRESSION OF STABLE ISOTOPE LABELLED PROTEIN WITHSILANTES RICH GROWTH MEDIA FOR YEAST OD 1 - 5

Silantes OD media are rich growth media for the expression of stable isotope labelled protein. The
media are available in all combinations of the stable isotopes 2H, 13C and 15N. They are formulated forincreasing cell densities (optical densities measured at 600nm) containing hydrolysates with increasing
concentrations. These OD-values are determined with common yeast stains such as P.pastoris.

The following instructions are applicable for

OD 1

OD 2OD 4OD 5OD 10The Silantes OD1-5 media are bacterial hydrolysates adjusted to M9 conditions. The glucose content
is less than 30mg/L.

Silantes OD1 media reach a cell density comparable to 0,2% glucose-M9 media.
Silantes OD2 media reach a cell density comparable to 0,4% glucose-M9 media.
Silantes OD2 media are commonly used if inducing the cells at OD0,6-0,9 followed by a ~4hincubation. The heterologous protein expression performance is similar as with LB medium.
Silantes OD4-, OD5- and OD10 (concentrate)-media are available on request.

The media are available as powder or as ready-to-use Silantes OD media solutions. If using the powder
media, proceed with the “Operating instructions for the preparation of Silantes OD Media Powder
for Yeast”. If using the ready-to-use media solution, proceed with the “Operating instructions for
Silantes OD Media Solutions for Yeast”. Please find both operating instructions on page 2.

This product is for laboratory use only. The safety and efficacy of this product in diagnostic or other clinical uses
is not established.

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MANUAL
PAGE 2

EXPRESSION OF STABLE ISOTOPE LABELLED PROTEIN WITHSILANTES RICH GROWTH MEDIA FOR YEAST OD 1 - 5

Operating instructions for the preparation of Silantes OD Media Powder for Yeast
Following instructions are applicable to obtain 1L of Silantes OD Media Solution. In order to avoid
isotopic dilution, use deuterated media powder (which is lyophilized from D2O-buffer) when preparing
the media solution in D2O.
1. Prepare 1000mL of H2O / D2O.
2. Add 10g Silantes Powder Media for Yeast.
3. Dissolve the powder.
The final yield cell density can be increased by 10-20%
by adding a salt mix at this stage.
4. Sterile filtrate the solution in an appropriate shaker bottle.

Salt mix and D2O
can be purchasedfrom Silantes.

After preparing the Silantes OD media solution for Yeast, proceed as follows:

Operating instructions for Silantes OD1-5 Media Solutions for Yeast
In order to avoid isotopic dilution by inoculation use an isotopically labelled Silantes OD-media as apreculture.
5. Fill Silantes OD into a sterile culture flask or fermenter.
For optimal aeration, fill your flask to a maximum of 1/3 of the flask volume.
6. If applicable for your system, add antibiotics and special nutrients.
7. Inoculate the Yeast culture.
8. Grow the Yeast culture in an orbital shaker or fermenter until the desired optical density isreached.
Ensure sufficient aeration. For optimal aeration, Silantes suggests to use an orbital shaker
apparatus at > 150 rpm.
9. Induce the expression of your protein in the usual way.
10. Collect the cells.

This product is for laboratory use only. The safety and efficacy of this product in diagnostic or other clinical uses
is not established.

www.silantes.com|

www.silantes-shop.com|

0049 (0)89 500 941 0|

info@silantes.com