Cell Signaling Technology Logo
1% for the planet logo

PathScan® Sandwich ELISA Lysis Buffer (1X) #7018

    Product Information

    Product Usage Information

    1. Thaw 10x buffer at 24-30°C, mixing end-over-end.

    2. Dilute 10X ELISA Lysis Buffer to a 1X solution using ddH2O.

    3. Chill 1X buffer on ice and add 1mM PMSF just prior to use.

    Lysis:

    All reagents and lysates must be kept cold.

    1. Treat cells as desired.

    2. Wash plate with PBS to remove residual media.

    3. Add 1X to cells:

    (a) For adherent cells, add 500-1000 µl of 1X ELISA Lysis buffer/10 cm dish.

    (b) For non-adherent cells, judgment must be used as to amount of lysis buffer to add. Addition of volume equal to cell pellet is generally appropriate

    4. Incubate plate on ice for 2 minutes.

    5. Collect cell lysate solution for use.

    6. Centrifuge extract for 5 minutes at 14,000 x g in a cold microfuge.

    7. Remove supernatant for use.

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

    Storage

    1X ELISA Lysis Buffer:

    20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM Na2 EDTA, 1 mM EGTA, 1% Triton, 20 mM sodium Pyrophosphate, 25 mM Sodium Fluoride, 1 mM β-glycerophosphate, 1 mM Na3VO4, 1 µg/ml leupeptin

    For long term storage, keep 10X buffer at -20°C If buffer will be continually used, it is recommended that the 10X buffer be kept at 4°C for 1-2 weeks. Aliquoting of 10X buffer is recommended if many small experiments are to be performed. 1X solution can also be aliquotted into single use aliquots and stored at -20°C.

    Product Description

    PathScan® Sandwich ELISA Lysis buffer is used to create cell extract for ELISA applications.

    Background

    PathScan® Sandwich ELISA Lysis buffer (1X) was developed for ELISA applications and is formulated to better inhibit Serine / Threonine class of phosphatases after cell extracts are made.
      For Research Use Only. Not for Use in Diagnostic Procedures.
      Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
      PathScan is a registered trademark of Cell Signaling Technology, Inc.
      All other trademarks are the property of their respective owners. Visit our Trademark Information page.