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Finest results are acquired when restricting extended cell exposure to ambient temperature disorders. Think about holding unused cells inside a humidified incubator with 5% CO2 at 37°C when accomplishing much larger experiments.
Evaluate the probable harmful results of prospect therapeutics, including small molecule compounds and biologics.
Get the job done swiftly at this stage to pool wells into a 15 mL tube. Note: For a large-scale dissociation, use a multichannel pipette to pool cells into a sterile reagent reservoir. Include the pooled wells into a 15 mL tube.
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Set the “unstained” tube aside. For the “viability dye” sample, centrifuge the tube at 300 x g
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In the event that cell clumping is observed immediately after thawing cryopreserved cells, it is recommended to filter aggregated suspensions by way of a 37 µm cell strainer (e.g. Catalog #27250) for optimal benefits. In such a case, the run time to the CellPore™ Transfection System might must be amplified to 10 seconds.
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When incorporating GCDR straight to the cultures, pipette up and down many periods to mechanically break up the dome totally making sure that GCDR can act on cells for dissociation.
Certainly, you’ll discover the action-by-stage protocol for TEER measurement To guage the epithelial barrier integrity in ALI cultures in this article.
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