Nutragen®

Note:Use these recommendations as guidelines to determine the optimal coating conditions for your culture system.

1. Remove required quantity of collagen from the bottle and dispense into a dilution vessel.
2. Dilute Nutragen®in water to ~50 to 100 µg/ml (~1:30). A 0.01 N HCl solution may also be used.
3. Swirl contents gently until material is completely mixed.
4. Add appropriate amount of diluted Nutragen® material to the culture surface ensuring that the entire surface is coated.
5. Incubate at room temperature or 37°C, covered, for 1-2 hours.
6. After incubation, aspirate any remaining material.
7. Rinse coated surfaces carefully with sterile medium or PBS, avoid scratching surfaces.
8. Coated surfaces are ready for use. They may also be stored at 2-8°C damp or air dried if sterility is maintained.

1. Slowly add 1 part of chilled 10X PBS or 10X culture media to 8 parts of chilled collagen solution with gentle swirling.
2. Adjust pH of mixture to 7.2–7.6. Use of sterile 0.1 M NaOH is recommended. Monitor pH adjustment carefully (pH meter, phenol red, or pH paper).
3. Adjust final volume to a total of 10 parts with sterile water.
4. To prevent gelation, maintain temperature of mixture at 2-10°C.
5. To form gel, warm to 37°C. For best results allow approximately 90 to 120 minutes for gel formation.