Whole Cell Lysate Preparation and Fractionation

For preparing whole cell lysate, HSC3 and HSC4 cells were washed twice with cold PBS, harvested and lysed with 0.1% SDS-added RIPA-buffer (Thermo Fisher Scientific, Tokyo, Japan) [32 (link)]. Cell fractions were extracted by using a Cell Fractionation Kit (Abcam, Cambridge, MA, USA), according to the manufacturer’s instructions [36 (link)]. Protein assay was performed using a Protein Assay Rapid Kit (Wako Pure Chemical Corporation, Osaka, Japan).

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Nakashima C., Yamamoto K., Kishi S., Sasaki T., Ohmori H., Fujiwara-Tani R., Mori S., Kawahara I., Nishiguchi Y., Mori T., Kondoh M., Luo Y., Kirita T, & Kuniyasu H. (2020). Clostridium perfringens enterotoxin induces claudin-4 to activate YAP in oral squamous cell carcinomas. Oncotarget, 11(4), 309-321.

Publication 2020

RIPA-buffer Cell Fractionation Kit Protein Assay Rapid Kit

A protein Assay Buffer Cell Cell fractionation Cold Protein Ripa

Corresponding Organization :

Other organizations : Nara Medical University, Osaka University, Nantong University

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Variable analysis

independent variables

Cell line (HSC3 and HSC4)

dependent variables

Whole cell lysate
Cell fractions

control variables

Washing cells twice with cold PBS
Lysis of cells with 0.1% SDS-added RIPA-buffer
Cell fractionation using Cell Fractionation Kit
Protein assay using Protein Assay Rapid Kit

controls

No positive or negative controls were explicitly mentioned.

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