IP Lysis Buffer for Animal Plant Cell, Fungal Bacterial Samples

Product Introduction

IP Lysis Buffer is a lysate that lyses cells or tissues to prepare protein samples under non-denaturing conditions. Protein samples obtained from tissue or cell lysis with this lysate can be applied to PAGE, western blot and immunoprecipitation (IP), co-immunoprecipitation (co-IP), Chromatin Immunopre-cipitation (ChIP) and ELISA where protein activity is required.

This product consists of 25 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA and 1% NP-40 can be applied to animal or plant tissue and cell samples, as well as for fungal or bacterial samples.

Storage

Ship with wet ice; Store away from light at 2-8℃, valid for 12 months.

Operation Step

Requires self-provide protease inhibitors. Protease inhibitors (Y20114, Y20115, Y20116, etc.) should be added to IP lysate before use to prevent protein degradation. All references to IP lysates in the following instructions have been added with protease inhibitors.

For tissue samples:

1.The tissue blocks are washed with pre-cooled PBS to remove blood stains, and then cut into small pieces and placed in the homogenizer.

2. Add 10 times tissue volume of IP Lysis Buffer and homogenize at low temperature (It is recommended to use high-speed tissue grinding instrument YB-HV, which is independently developed and produced by Yanbiotech). Note that the amount of IP Lysis Buffer can be added at a ratio of approximately 50 mg of tissue to 1 mL of lysate. If the tissue protein content is low, the amount of lysate can be reduced to increase the protein concentration in the crude extract solution.

3. Transfer the homogenate to a 1.5 mL of centrifuge tube and shake. Ice bath for 30 min with repeated pipetting every 10 min to ensure complete lysis of tissue cells;

4. Centrifuge at 12000 x g for 5 min and collect the supernatant, as total protein solution.

For adherent cell samples:

1. Wash the cells with PBS 2-3 times, and aspirate the residual liquid thoroughly at the last time.

2. Absorb IP Lysis Buffer into the cell culture plate and flask at the ratio of 250 μL of lysate per well of the 6-well plate, shake the culture plate and flask repeatedly to make the lysate fully contact with the cells for 3-5 min.

3. Scrape off the cells with a cell scraper and collect them into a centrifuge tube.

4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

For suspended cell samples:

1. Cells were collected by centrifugation.

2. Mix the cytosol with IP lysate at a ratio of 250 μL of lysate per cell well of a 6-well plate, and shake.

3. Ice bath for 30 min, repeatedly pipetting every 10 min to ensure complete cell lysis.

4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

For bacterial or fungal samples:

1. 1 mL of the bacterial suspension was centrifuged to remove the supernatant and washed once with PBS to fully remove the liquid. The vortices make the thalli disperse as much as possible.

2. Add 100-200 μL of IP lysate and vortex gently to mix well with the lysate.

3. Ice bath for 10 min, repeatedly pipetting every 2 min to ensure complete lysis of thalli.

4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

Notice

1. Tissue or cells may appear viscous when lysed. This can be achieved by repeatedly blowing with a pipettor or shaking with a vortexer until it becomes a liquid. Add a further appropriate amount of lysis solution if it remains thick.

2. This reagent does not contain protease inhibitors, Add your own protease inhibitors before use. Y20114, Y20115, Y20116 and other related protease inhibitors of our company are recommended.

3. The total protein solution obtained by cleavage of this product is compatible with the BCA protein quantitative detection kit (Y20122) of our company.

4. For your safety and health, please wear safety glasses, gloves, or protective clothing.