News from the Bench
Discover what’s going on at CST, receive our latest application notes and tips, read our science features, and learn about our products.
Our WB Protocol, now in a kit.
To Purchase # 9803S
|$ 0. 00|
Find answers on our FAQs page.
PTM information and tools available.
Cell Lysis Buffer (10X) #9803
Gallery: Cell Lysis Buffer (10X) #9803
Cell Lysis Buffer is used to lyse cells under nondenaturing conditions.
1. If buffer will be continually used, it is recommended that the 10x buffer be kept at 4°C for 1-2 weeks. For longer periods of time, buffer should be stored at –20°C. Aliquotting of 10x buffer is recommended if many small experiments are to be performed.
2. Thaw 10x buffer at 24-30°C, mixing end-over-end.
3. Dilute 10X Cell Lysis Buffer to a 1X solution using ddH2O. This product supplies enough 10X material to make 150mls of whole cell extract.
4. Chill 1X buffer on ice and add PMSF just prior to use.
Note: CST recommends adding 1 mM PMSF immediately before use.
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 lysis buffer/ 10 cm dish.
4. Incubate plate on ice for 5 minutes.
5. Scrape cells.
6. Sonicate briefly.
7. Spin extract 10 minutes at 14,000 x g in a cold microfuge.
8. Remove supernant for use.
1. For non-adherent cells, add 400 µl of buffer per 107 cells once they have been washed in 1X PBS and pelleted.
2. 2X #9803 Cell Lysis 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.Solutions and Reagents:
20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM Na2EDTA, 1 mM EGTA, 1% Triton, 2.5 mM sodium pyrophosphate, 1 mM beta-glycerophosphate, 1 mM Na3VO4, 1 µg/ml leupeptin
Note: CST recommends adding 1 mM PMSF immediately before use.Storage: Store at -20°C. For short term storage (1-2 weeks), Cell Lysis Buffer can be stored at 4°C.
For Research Use Only. Not For Use In Diagnostic Procedures. Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.