RIPA Lysis Buffer System

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Cells are thawed on ice and resuspended in 3-5X packed-cell volume of ice-cold RIPA Lysis Buffer System according to the manufacturer’s instructions and freshly supplemented with 1X PhosSTOP phosphatase inhibitor cocktail when applicable.	- Cell pellets are homogenized by 5-10 passages through a 21-gauge needle then mixed for 60 minutes at 4 °C on an automatic rotator. Insoluble cellular debris is pelleted using centrifugation at 13000 rpm for 10 mins at 4 °C.

Protocol tips
- Cells are thawed on ice and resuspended in 3-5X packed-cell volume of ice-cold RIPA Lysis Buffer System according to the manufacturer’s instructions and freshly supplemented with 1X PhosSTOP phosphatase inhibitor cocktail when applicable.
Downstream tips
- Cell pellets are homogenized by 5-10 passages through a 21-gauge needle then mixed for 60 minutes at 4 °C on an automatic rotator. Insoluble cellular debris is pelleted using centrifugation at 13000 rpm for 10 mins at 4 °C.

Publication protocol

Cells were thawed on ice and resuspended in 3-5X packed-cell volume of ice-cold RIPA Lysis Buffer System (Santa Cruz Biotechnology, Dallas, TX, USA) according to the manufacturer’s instructions and freshly supplemented with 1X PhosSTOP
phosphatase inhibitor cocktail when applicable (Roche [Mississauga, ON, Canada]). Cell pellets were homogenized by 5-10 passages through a 21-gauge needle then mixed for 60 minutes at 4 °C on an automatic rotator. Insoluble cellular debris was pelleted using centrifugation at 13000 rpm for 10 mins at 4 °C.

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Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for RIPA Lysis Buffer System below.

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