WB Lysis and Extraction Buffer

Overview

Tested applications

Unless otherwise stated in our catalog or other company documentation accompanying the product, our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Properties

Form

Liquid

Ready to use.

Storage instruction

Store at 4°C

Applications

Protocols

Lysis of Mammalian Whole Cell:

 

For lysis of adherent cells, we recommend the following: (all reagents and lysates must be kept cold).

 

1. Treat cells as desired.

 

2. Wash plate with PBS to remove residual media.

 

3. Add 400 µl of 1x buffer/10 cm dish.

 

4. Incubate plate on ice for 5 minutes.

 

5. Scrape cells.

 

6. Sonicate briefly.

 

7. Centrifuge extract for 10 minutes at 14,000 x g in a cold microfuge.

 

8. Remove supernatant for use.

 

Additional notes:

 

1. For non-adherent cells, add 400 µl of buffer per 107cells once they have been washed in 1X PBS and pelleted.

 

2. 1X Buffer can be used for lysis of tissue samples, although a homogenization step is recommended after adding lysis buffer. Extract the tissue at a ratio of 100 mg of tissue to 1 ml of buffer. Sonication of the tissue lysate is also required.

 

3. Additional protease inhibitors can be added to the 1x lysis buffer without any difficulties.