RIPA Lysis Buffer, 10X

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Cells are lysed with RIPA lysis buffer containing Halt™ Protease Inhibitor Cocktail.

Protocol tips
- Cells are lysed with RIPA lysis buffer containing Halt™ Protease Inhibitor Cocktail.

Publication protocol

HaCaT were trypsinized and washed with PBS buffer twice. Cells were lysed with RIPA lysis buffer (Merck, Germany) containing Halt™ Protease Inhibitor Cocktail (Thermo Scientific, USA) and protein concentration was quantified BCA assay (Thermo Scientific). A total of 20 μg (GAPDH) and 60 μg (CASP3) proteins were separately loaded onto 12% SDS-PAGE gels and were transferred onto a polyvinylidene difluoride membrane (PVDF; GE Healthcare, UK). The membrane proteins were immunoblotted with primary mAb . After washing, the protein membrane of was incubated with anti-mouse horseradish peroxidase-conjugated secondary antibody. The signals were analyzed with enhanced SuperSignal™ West Femto Maximum Sensitivity Substrate (Thermo
Scientific). Images were quantified using a Licor-odyssey.

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