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Specification Sheet: Nunc 300 cm2 Cell Culture Flask Product Literature

Technical Note: Culturing technique for Nunc TripleFlask Treated Cell Culture Flasks Product Literature

How do I process cell culture supernatants for use on the Luminex assay platform? Product FAQ

Answer

Cells should be in log-phase growth. Stimulate cells as desired in appropriate cell culture flasks. Using sterile technique, remove the desired volume of conditioned cell culture medium with a pipette and transfer the medium to clean polypropylene microcentrifuge tubes. Centrifuge the medium at 14,000 rpm for 10 min at 4 degrees C in a refrigerated microcentrifuge to remove any cells or cellular debris. Aliquot the clarified medium into clean polypropylene microcentrifuge tubes. These samples are ready for the assay. Alternatively, clarified medium samples can be aliquoted and stored at -80 degrees C for future analysis. Avoid multiple freeze-thaw cycles. Frozen samples should be allowed to thaw on ice just prior to running the assay. Thawed samples should be clarified by centrifuging at 14,000 rpm for 10 min at 4 degrees C in a refrigerated microcentrifuge prior to analysis to prevent clogging of the Luminex probe and/or filter plate. Follow the assay procedure provided with the kit for appropriate dilutions.

Answer Id:: E12642

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Smart Note: How do I evenly distribute my cell suspension among the multiple growth surfaces in a triple-layer flask? Product Literature

Can I use a Nunc Cell Scraper with a Nunc TripleFlask that has Nunclon Delta surface treatment? Product FAQ

Answer

The design of the Nunc TripleFlask does not allow for mechanical scraping. However, cells are easily harvested using protein digesting enzymes, such as trypsin, collagenase, and Pronase or by using non-enzymatic dissociation solutions.
The following is a document link regarding culturing techniques for Nunc TripleFlask:
https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/culturing-technique-nunc-tripleflask-technical-note.pdf

Answer Id:: E17707

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What is the recommended working volume of Nunc adherent cell culture products? Product FAQ

Answer

This depends largely on the cell line and working conditions (considerations include headspace for gas exchange, supplements used, and optimal feeding schedule, among others) but we do offer guidelines for development purposes (0.3 to 0.5 mL per cm2) . Note that these volumes are approximate and may have to be modified for best performance. T25 Flask = 7 mL, T75 Flask = 25 mL, T175 Flask = 55 mL, T225 Flask = 70 mL, 300cm2 Flask = 100 mL, 500 cm2 TripleFlask = 200 mL, 35 mm round culture dish = 3 mL, 60 mm round culture dish = 5 mL, 100 mm round culture dish = 12 mL, 150 mm round culture dish = 35 mL, 4 (rectangular) well Multidish = 5 mL, 8 (rectangular) well Multidish = 3 mL, 4 (round) well Multidish = 1 mL, 6 well Multidish = 3 mL, 12 well Multidish = 2 mL, 24 well Multidish = 1 mL, 48 well Multidish = 0.5 mL.

Answer Id:: E15931

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