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9803
Cell Lysis Buffer (10X)
Buffers & Dyes

Cell Lysis Buffer (10X) #9803

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Product Usage Information

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.

Lysis:

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.

Additional notes:

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:

This product is stable for 24 months when stored at -20°C. Cell Lysis Buffer can be stored at 4°C for a short period of time (1-2 weeks).

Product Description

Cell Lysis Buffer is used to lyse cells under nondenaturing conditions.

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

To Purchase # 9803S
Product # Size Price
9803S
15 ml N/A