CelLytic™ M

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	- Protocol was performed following the manufacturer's instructions.

Protocol tips
- Protocol was performed following the manufacturer's instructions.

Publication protocol

Cells were lysed in Cell Lytic M (C2978, Sigma-Aldrich, San Luis, MS, USA) plus protease inhibitors (11836145001, Roche, Basel, Switzerland). Protein concentrations were determined by Bradford assay (23200, Thermo Fischer Scientific, Waltham, MA, USA) using a Nanodrop™ 2000 spectrophotometer (ND-2000, Thermo Fischer Scientific, Waltham, MA, USA). 1.5 µg of protein was solubilized with 6× DTT sample buffer (Tris-HCl, Glycerin, SDS, DTT, Bromophenol), boiled for 5 min at 95 °C, cooled down and resolved by electrophoresis. Proteins were blotted to a PVDF membrane (GE Healthcare, Braunschweig, Germany) by Mini Trans-Blot® (Bio Rad, Hercules, CA, USA) wet transfer system. Membranes were blocked and incubated with ab in 5% milk powder with 0.1% PBS-Tween20 either for 1 h at RT or o/n at 4 °C with constant shaking. Protein was detected using the HRP-based ECL chemiluminescence system (GE Healthcare, Braunschweig, Germany) and blots were documented with a LAS-4000 Mini (Fujifilm, Düsseldorf, Germany).

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for CelLytic™ M below.

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