How should I reconstitute Efficient-Pro AGT Medium (Cat. No. A5322301, A5322302, A5322303, A5322304)?
Please follow the steps shown below to reconstitute Efficient-Pro AGT Medium (Cat. No. A5322301, A5322302, A5322303, A5322304) and to make Complete Medium:
- Measure 90% of the final volume of deionized or distilled water at room temperature (15-30 degrees C).
- Add Efficient-Pro AGT Medium at 27.3 grams/L to water.
- Mix for a minimum of 20 min.
- Use a calibrated vessel to dilute to final production volume with ambient deionized or distilled water.
- Mix for an additional 20 min.
- Measure the pH, then measure and record osmolality.
- Sterilize the product immediately by membrane filtration(positive pressure recommended).
Note: Once the product is filtered, use immediately or store at 2-8 degrees C for up to 6 months. Protect from light.
To make Complete Medium,
- Supplement Efficient-Pro AGT Medium with L‑glutamine or GlutaMAX Supplement at 2-8 mM final concentration prior to use (except for CHO-K1 GS cell lines).
Note: The recommended L‑glutamine and/or GlutaMAX Supplement concentration is 6 mM.
- Add 10 mL/L of HT Supplement for use in applications not requiring DHFR amplification.
- Glucose supplementation may be required for terminal batch cultures and should be determined empirically.
- Add Anti-Clumping Agent (1 mL/L) to the medium to reduce cell aggregation, if required.
Note: Consider reducing L‑glutamine concentration for fed-batch or perfusion protocols, or if the cell line in-use is sensitive to ammonia. Addition of a surfactant such as Pluronic™ F-68 is not required.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
What is the shelf-life of Efficient-Pro AGT Medium?
The shelf-life of Efficient-Pro AGT Medium is 12 months from the date of manufacture. The expiration date is provided on the product label.
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If my cell line is sensitive to ammonia, can I still use Efficient-Pro AGT Medium?
Yes, however, we recommend reducing the L-glutamine concentration if a decrease in cell viability is observed. The recommended glutamine concentration is 2-8 mM. Another way to reduced ammonia production is to use GlutaMAX Supplement instead of L-glutamine. Addition of Pluronic F-68 is not required.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Does Efficient-Pro AGT Medium require supplementation for use?
When using a CHO cell line with a selection system other than the glutamine synthetase (GS) system, we recommend adding 2-8 mM L-glutamine or GlutaMAX Supplement to Efficient-Pro AGT Medium. Additionally, we recommend making sure that glucose levels do not fall below 2 g/L in culture by supplementing with glucose as needed. Also, if clumping becomes an issue, then add anti-clumping agent (ACA) to your cultures at 1% v/v.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
What is the concentration of glucose in Efficient Pro-AGT Medium?
Efficient Pro-AGT Medium contains 4.7 g/L of glucose.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.