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WB/IP Lysis Buffer

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MCE WB/IP Lysis Buffer is a lysis buffer used to lyse cell or tissue samples under non-denaturing conditions to prepare protein samples. The lysed cell or tissue samples can be used for PAGE, Western Blot, Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), ELISA, and other assays.

  • Storage :
    -20℃,1 year.
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Description & Advantages

MCE WB/IP Lysis Buffer is a lysis buffer used to lyse cell or tissue samples under non-denaturing conditions to prepare protein samples. The lysed cell or tissue samples can be used for PAGE, Western Blot, Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), ELISA, and other assays.

This product contains various inhibitors such as sodium pyrophosphate, β-glycerophosphate, sodium orthovanadate, EDTA, and leupeptin, which effectively prevent protein degradation and maintain the original protein-protein interactions.

Protocol

General Protocol

Experiment Preparation

Take an appropriate amount of WB/IP Lysis Buffer and add PMSF, protease inhibitor, or phosphatase inhibitor according to experimental needs. Mix and keep on ice.

1. Cell protein extraction

Adherent cells:

1) Carefully remove culture medium and wash once with PBS, saline, or serum-free medium. Washing may be omitted if serum proteins do not interfere with downstream experiments.

2) Add 100-200 μL lysis buffer per well of a 6-well plate and pipette several times to ensure full contact with the cells.

Suspension cells:

1) Collect cells by centrifugation at 2,500 × g for 10 min at 4°C and gently tap the tube to loosen the pellet.

2) Add lysis buffer at an amount equivalent to 100-200 μL per well of a 6-well plate. For large cell numbers, divide into 5 × 105-1 × 106 cells/tube before lysis.

Recommended lysis volumes for different culture vessels:

Culture vessel Lysis Buffer volume
100 mm dish 500-1000 μL
60 mm dish 200-400 μL
6-well 100-200 μL
12-well 50-100 μL
24-well 25-50 μL
48-well 12.5-25 μL
96-well 5-10 μL

3) Incubate the sample on ice for 2-10 min for complete cell lysis.

4) Collect the lysate and centrifuge at 14,000 × g for 3-5 min at 4°C.

5) Transfer the supernatant to a new tube for PAGE, Western Blot, IP, Co-IP, or other downstream analyses.

2. Tissue protein extraction

1) Quickly cut tissue into small pieces or freeze in liquid nitrogen and grind into powder.

2) Add 100-200 μL WB/IP Lysis Buffer per 20 mg tissue, homogenize thoroughly, and incubate on ice for 2-10 min.

3) Centrifuge at 14,000 × g for 3-5 min at 4°C.

4) Transfer the supernatant to a new tube for PAGE, Western Blot, IP, Co-IP, or other downstream analyses.

Note: Perform all protein extraction steps on ice or at 4°C whenever possible. Because of the detergent concentration, Bradford assays are not recommended; BCA assays may be used for protein quantification.

Storage

-20℃,1 year.

Components

Components HY-K1000-100 mL
WB/IP Lysis Buffer 100 mL

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