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NAT2 in-cell western staining in cryoplateable human hepatocytes

RS Raúl A. Salazar-González
MD Mark A. Doll
DH David W. Hein
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Cryoplateable human hepatocytes (2×105 viable cells) were plated into 96-well black/clear bottom plates (Thermo Fisher Scientific, Waltham, MA, USA) and incubated overnight at 37 °C and 5% CO2 on InVitroGRO HT medium (BioIVT, Baltimore, MD, USA). Once attached to the plate, cells were fixed, permeabilized and blocked following the same protocol previously described for UV5 cells. After blocking, cells were incubated with NAT2 specific antibody (ab194114 (1:200)), and β-actin (A2228 (1:200)), Sigma-Aldrich, St. Louis, MO, USA) overnight at 4 °C with constant agitation. Following primary antibodies incubation, the same protocol previously described for UV5 cells was followed for secondary antibodies incubation and detection.

Copyright and License information: The Author(s) ©2020
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