Product Composition

Product Introduction

The  RIPA  tissue/cell lysis buffer lyses cell membranes (including nuclear membranes) by means of surfactants and other components. The proteins extracted by this reagent are soluble total proteins.

Protocols(for reference only)

If precipitation is found in the RIPA lysis buffer, place it at room temperature for half an hour or in a room-temperature water bath to dissolve the precipitate.

According to the required volume, add 10 μL of PMSF to each 1 mL of RIPA lysis buffer to make the final concentration of PMSF 1 mM, and mix well for use (PMSF should be added fresh before use).

1. Sample Pre-treatment
For adherent cells: Remove the culture medium and rinse the cells once with PBS, normal saline or serum-free culture medium.  Add  the lysis buffer at a ratio of 150-250 μL per well of a 6-well plate. Pipette up and down several times to ensure full contact   between the  lysis buffer and the cells.
For suspension cells: Collect the cells by centrifugation and add the lysis buffer at a ratio of 150-250 μL per well of a 6-well plate.  Flick the tube containing the suspension cells gently with your fingers to achieve complete lysis. There should be no obvious cell  precipitation after sufficient lysis. If the cell amount is large, the cells must be aliquoted into tubes with 5- 10×10⁵ cells per tube  before lysis.
For tissue samples: Cut the tissue into small pieces. Add the lysis buffer at a ratio of 150-250 μL per 20 mg of Tissue (if lysis is      insufficient, an appropriate amount of additional lysis buffer can be added; if a high-concentration protein  sample is required, the amount of lysis buffer can be reduced appropriately). Homogenize with a glass homogenizer until complete lysis is achieved.

2. Post-treatment 
Centrifuge the lysed samples at 10,000- 14,000 g for 3-5 minutes, collect the supernatant, and proceed to subsequent  operations  such as protein concentration determination,SDS-PAGE, Western blotting and co-immunoprecipitation.