RIPA Cell lysis buffer

一. It is applicable to the lysis of tissues, cells, bacteria, etc., and protein interactions can be retained after lysis

二. Experimental Procedures

1. Place the culture flask filled with cells on ice and use a pipette to draw out the culture medium. Add sufficient cold PBS to the culture flask to thoroughly wash the cell surface to remove any remaining culture medium. Discard the PBS and repeat the above operation 2 to 3 times. In the last wash, absorb as much residual PBS as possible and operate on ice whenever possible.

2. Add cold RIPA lysis buffer to the culture flask (1ml for every 75cm ² of culture flask). Then, use a cell scraper to start scraping the cells along the bottle wall. If there are multiple bottles of the same type of cells to be scraped, draw out the cell fluid scraped from the first bottle and transfer it to the next bottle for further scraping (since the treated cell lysate is relatively viscous, it is advisable to use a larger diameter pipette for drawing).

3. Aspirate the scraped cell lysate and place it in a 14ml centrifuge tube (on ice), then repeat step 2 to scrape off the remaining cells.

4. Collect as much cell lysis buffer as possible into a 14ml centrifuge tube. Insert the sample into an ice box for ultrasound. The ultrasound intensity should be such that no foam is produced. Each ultrasound session should last for 2 to 3 seconds, and repeat 3 to 4 times. If there are still cell fragments or precipitates, centrifuge at 10,000 RPM for 10 minutes and retain the supernatant.

5. Take a small amount of the cell lysate to measure its protein concentration (using a Biorad Bradford kit ora UV spectrophotometer at A280), aliquot and store at -70℃.

Transportation and storage：Dry ice transportation, store at -20℃

Declaration: This product is for scientific research only.