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Best final results are acquired when limiting extended cell publicity to ambient temperature disorders. Think about retaining unused cells in the humidified incubator with five% CO2 at 37°C when accomplishing larger experiments.
Each one of these cells has a role to Participate in. Investigate on how they get the job done as well as their interactions with other subsets carries on to reveal opportunities for novel therapeutic tactics.
Utilizing a P1000 pipette, Carefully triturate BLOs within the apical chamber somewhere around 3 times even though circling the idea across the insert. Transfer contents to some fifteen mL conical tube.
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Established the “unstained” tube aside. For the “viability dye” sample, centrifuge the tube at three hundred x g
Heading again into the plate, rinse Each and every properly with one mL of FACS buffer and transfer the quantity towards the 15 mL tube. Notice: Keep cell suspension on ice soon after transfer to your tube right until prepared to operate FACS.
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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 this case, the run time to the CellPore™ Transfection System might have to be increased to ten seconds.
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Notes: Use pipette suggestions to Carefully scratch 줄기세포 지방이식 the bottom of each properly in order to lift the monolayer off, then dissociate the cell clumps into an individual-cell suspension by pipetting up and down five - 줄기세포 지방이식 ten situations.
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Take away supernatant and resuspend cells in FACS buffer. Be aware: It's important to quench the dissociation reagent by utilizing the identical or double the volume with the dissociation reagent.