Products
Protein Biology Reagents | Consumables
Protein Extraction
Lysis Buffer
Strong RIPA Lysis Buffer
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หมวดหมู่ : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
แบรนด์ : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
RIPA Lysis Buffer (Strong) | G2002-30ML | 30 mL |
G2002-100ML | 100 mL |
Product Description/Introduction
RIPA Lysis Buffer is a traditional rapid lysis solution for cells and tissues. It is available in a variety of formulations and classified as strong, medium or weak. The protein samples obtained from the lysis of tissues and cells in RIPA strong lysis solutions can be used for routine PAGE, Western and other experiments where protein activity is not strictly required. This product contains 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM EDTA-2Na, 1% Triton X-100, 1% sodium deoxycholate and 0.1% SDS. It is suitable for animal or plant tissues and cells, but can also be used for fungal or bacterial samples.
Storage and Shipping Conditions
Ship with wet ice; Store in the dark at 2-8; valid for 18 months.
Product Components
Component | G2002-30ML | G2002-100ML |
RIPA Lysis Buffer (Strong) | 30 mL | 100 mL |
Manual | 1 pc | |
Preparation for The Experiment
Requires self-provide protease inhibitors. Protease inhibitors (G2006, G2007, G2008, etc.) should be added to RIPA Lysis (Strong) before use to prevent protein degradation. All references to RIPA Lysate (Strong) in the following instructions have been added with protease inhibitors.
Assay Protocol/Procedures
l For tissue samples
1. The tissue pieces are washed with pre-cooled PBS (G4202 recommended) to remove blood stains, cut into small pieces and placed in the homogenizer.
2. Add 10 times the tissue volume of RIPA lysis buffer (strong) and homogenate at low temperature (It is recommended to use high-speed tissue grinding instrument KZ-III-F and KZ-III-FP, which is independently developed and produced by Servicebio). Note that the amount of RIPA lysate (strong) can be added in a ratio of approximately 50 mg of tissue to 1 mL of lysate. If the content of tissue protein is low, the amount of lysate can be reduced to increase the protein concentration in crude extract solution.
3. Transfer the homogenate to a 1.5 mL of centrifuge tube and oscillate. Ice bath for 30 min with repeated pipetting every 10 min to ensure the complete lysis of tissue cells.
4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.
l For adherent cell samples
1. Wash the cells with PBS for 2-3 times and aspirate the residual liquid thoroughly at the last time.
2. Aspirate RIPA Lysis Buffer (Strong) into the cell culture plate and flask at a ratio of 250 μL of cell lysate per well of 6-well plate, shake the culture plate and flask repeatedly to make the lysate fully contact with cells for 3-5 min.
3. Scrape off the cells with a cell scraper and collect into a centrifuge tube.
4. Lysis on ice for 30 min.
5. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.
l For suspension cell samples
1. Centrifuge.to collect cells.
2. Mix the cytosol with RIPA Lysis Buffer (Strong) 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.
l For bacterial or fungal samples
1. Remove 1 mL of the suspension, centrifuge to remove the supernatant and wash once with PBS to fully remove the liquid. Vortex to disperse the bacterium as much as possible.
2. Add 100-200 μL of RIPA Lysis Buffer (Strong) and vortex gently to mix well with the lysate.
3. Ice bath for 30 min, repeatedly pipetting every 2 min to ensure complete cell lysis.
4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.
Note
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. G2006, G2007, G2008 and other related protease inhibitors of our company are recommended.
3. For your safety and health, please wear safety glasses, gloves, or protective clothing.
For Research Use Only!
