HiCultS9 | cultivated, cell line-derived human S9 fraction

HiCultS9 is a first of its kind human in vitro metabolisation agent derived from a chemically defined, animal component- as well as protein-free production process.
It replaces the commonly used pooled human liver S9 fraction and can be used as alternative to the rat liver S9 fraction as well.

The supplement is available in two variations:

• As a frozen liquid (5 mg | 5 mg/ml)  (109101-5MG)
• As a lyophilised solid (5 mg)  (109102-5MG), which provides improved shipping and storage behaviour and is reconstituted in water

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Application note | Characteristics

HiCultS9 is sterile and ready-to-use upon reconstitution or thawing, respectively. This metabolic agent can by applied such as standard human S9 or its animal variants. The usual concentration is 0.1 - 1.0 mg/ml within your buffered cofactor mix consiting of NADP, glucose-6-phosphate and magnesium salts. The enzyme activity for NADPH regeneration is sufficient for standard applications.

Since it is sterility tested and free of mycoplasma, HiCultS9 can be applied to serum-free as well as FCS-containing cell cultures directly at a concentration of 50 - 500 mg/L final volume.

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Protocol

Standard metabolisation

• Dissolve your compounds in sodium phosphate buffer (pH 7.4; 100 mM);
• Add the same voulme of a mixture of 0.2 - 2.0 mg/ml HiCultS9 in sodium phosphate buffer (pH 7.4; 100 mM) containing 6 mM NADP, 30 mM glucose-6-phosphate and 10 mM magnesium chloride;
• Incubate the mixture for 30-120 minutes at 33 - 37 °C under constant shaking.