Ature Transfer supernatant cautiously with 25 mL Pipette to 50 mL conical, discard pelletEur J Immunol. Author manuscript; available in PMC 2020 July 10.Cossarizza et al.PageFill up to 50 mL with PBS/Hank’s Centrifuge 4 min/40 g/room IL-17C Proteins Source nitrogendDependingeAdheringto standard density centrifugation protocol is relevant for this step. Use minimum acceleration and no brakes on the centrifuge. preservation of isolate leukocytes may be performed at this step (see also d): Centrifuge five min/500 g/room temperature discard supernatant Resuspend pellet in freezing medium to get a final concentration of 1 107 cells/mL Pipette cells into cryo tubes (1 mL every single) Put tubes into precooled stratacooler and retailer at -80 for 24 h prior to transferring into liquid nitrogenfCyrogCellsstored at these points have already been successfully utilised for each phenotypical as well as functional evaluation. When making use of cells stored without having leukocyte purification s.
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