• PN: P820650

CONTENT: One bottle containing 5 mL of 10x Tissue Lysis Buffer

STORAGE:  -20 C.

APPLICATION: Optimal assay component for AimPlex immunoassays. For preparing tissue or cell lysates for AimPlex immunoassays by following the procedures described below. Refer to the AimPlex Multiplex Immunoassay User Manual and kit inserts for the assay procedure after the lysates are prepared.

PREPARING 1x Tissue Lysis Buffer: Thaw the 10x Tissue Lysis Buffer on ice before use. Mix 5 mL of the 10x Tissue Lysis Buffer with 45 mL ddH2O. Note: Additional protease inhibitors such as PMSF (at a final concentration of 1mM) can be added to the lysis buffer right before use.

PREPARING LYSATE FROM ADHERENT CELLS: 1. Seed, grow and treat cells under appropriate experimental conditions. 2. Remove culture medium by aspiration. 3. Wash once with ice cold 1x PBS (Optional). 4. Add 200 µL of ice cold lysis buffer/well of a 24-well plate. 5. Pipette up and down 10-12 times . 6. Incubate on ice for 5 min. 7. Centrifuge extract at 14,000xg for 10 minutes at 2-8 oC. 8. Collect the supernatant fraction for AimPlex assays.

PREPARING LYSATE FROM NON-ADHERENT CELLS: 1. Seed, grow and treat cells under appropriate experimental conditions. 2. Pellet cells by centrifugation (300xg, 5 minutes at 2-8 oC). 3. Wash once with ice cold 1x PBS (Optional). 4. Add 500 µL of ice cold lysis buffer/107 cells. 5. Pipette up and down 10-12 times . 6. Incubate on ice for 5 min. 7. Centrifuge extract at 14,000 x g for 10 minutes at 2-8 oC. 8. Collect the supernatant fraction for AimPlex assays.

PREPARING TISSUE LYSATE: 1. Add 500 µL of ice cold lysis buffer/100 mg of tissue. 2. Homogenize at 2-8 oC by using a tissue homogenizer such as POLYTRON* PT1300D or QIAGEN's TissueLyser*. 3. Incubate on ice for 5 min. 4. Centrifuge extract at 14,000 x g for 10 minutes at 2-8 oC. 5. Collect the supernatant fraction for AimPlex assays.

NOTE: If measuring protein concentration in the lysate is needed, Lowry Protein Assay is recommended because the lysis buffer contains Triton* X-100

For Research Use Only.  Not for use in diagnostic procedures.

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