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XML generation date: 2024-10-17 20:56:39.400
Product last modified at: 2024-10-23T23:15:08.498Z
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PDP - Template Name: Buffer
PDP - Template ID: *******ceb613c

Cell Lysis Buffer (10X) #9803

    Product Information

    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. Aliquoting 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 supernatant 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.

    Storage

    1X Cell Lysis Buffer:

    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.

    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.
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