5X RIPA Buffer Reagent
5X RIPA Buffer
Synonyms
5X RIPA Buffer; 5X RIPA Buffer reagent
Preparation of the Reagents
Mix the 5X RIPA Buffer by inverting/vortexing the bottle a few minutes. Dilute the 5X RIPA Buffer to 1X with deionized water. Stir to homogeneity. Store at 4 degree C for up to 6 months.
Quantity
20 mL
Preparation of Samples
I. Adherent Cells
1. Culture cells to approximately 80-90% confluence.
2. Aspirate the culture media and wash twice with cold PBS.
3. Add cold 1X RIPA Buffer to the cells, spreading evenly. Use 1 mL of buffer per 10-cm dish (56.7 cm2), maintaining on ice for 5 minutes. Cells may be scraped with a cell scraper. 4. Collect the lysate into a microcentrifuge tube.
5. Centrifuge for 10 minutes (14000 x g).
6. Transfer the supernatant to a clean tube and maintain at 4 degree C. Samples may be frozen at -80 degree C.
II. Suspension Cells 1. Collect the cells into an appropriate conical centrifuge tube.
2. Centrifuge for 5 minutes (600 x g).
3. Remove and discard the supernatant.
4. Wash the cells twice with cold PBS, centrifuging for 5 minutes between (600 x g).
5. Add cold 1X RIPA Buffer to the cell pellet. Use 1 mL of buffer per 40 mg of wet cell pellet. Gently resuspend the pellet by pipetting up and down.
6. Shake the suspension for 15 minutes at 4 degree C. 7. Centrifuge for 10 minutes (14000 x g).
8. Transfer the supernatant to a clean tube and maintain at 4 degree C. Samples may be frozen at -80 degree C.
1. Culture cells to approximately 80-90% confluence.
2. Aspirate the culture media and wash twice with cold PBS.
3. Add cold 1X RIPA Buffer to the cells, spreading evenly. Use 1 mL of buffer per 10-cm dish (56.7 cm2), maintaining on ice for 5 minutes. Cells may be scraped with a cell scraper. 4. Collect the lysate into a microcentrifuge tube.
5. Centrifuge for 10 minutes (14000 x g).
6. Transfer the supernatant to a clean tube and maintain at 4 degree C. Samples may be frozen at -80 degree C.
II. Suspension Cells 1. Collect the cells into an appropriate conical centrifuge tube.
2. Centrifuge for 5 minutes (600 x g).
3. Remove and discard the supernatant.
4. Wash the cells twice with cold PBS, centrifuging for 5 minutes between (600 x g).
5. Add cold 1X RIPA Buffer to the cell pellet. Use 1 mL of buffer per 40 mg of wet cell pellet. Gently resuspend the pellet by pipetting up and down.
6. Shake the suspension for 15 minutes at 4 degree C. 7. Centrifuge for 10 minutes (14000 x g).
8. Transfer the supernatant to a clean tube and maintain at 4 degree C. Samples may be frozen at -80 degree C.
Preparation and Storage
Store RIPA Buffer concentrate at room temperature.. Shelf Life: 1 year from date of receipt under proper storage conditions; aliquot to avoid multiple freeze thaw cycles
Related Product Information for 5X RIPA Buffer reagent
• Buffer is able to extract cytoplasmic, membrane, and nuclear proteins
• Suitable for downstream assays such as reporter assays, protein assays, immunoassays and protein purification
RIPA Buffer continues to be a popular buffer to lyse plated and suspension cultured mammalian cells. The buffer extracts cytoplasmic, membrane, and nuclear proteins and is suitable for downstream assays such as reporter assays, protein assays, immunoassays and protein purification.
• Suitable for downstream assays such as reporter assays, protein assays, immunoassays and protein purification
RIPA Buffer continues to be a popular buffer to lyse plated and suspension cultured mammalian cells. The buffer extracts cytoplasmic, membrane, and nuclear proteins and is suitable for downstream assays such as reporter assays, protein assays, immunoassays and protein purification.
Product Categories/Family for 5X RIPA Buffer reagent
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Product Notes
The 5X RIPA Buffer (Catalog #AAA168128) is a Reagent and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "5X RIPA Buffer, Reagent" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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