Get Ripa Buffer (10x) - Cell Signaling Technology

This guide provides clear and comprehensive instructions for filling out the RIPA Buffer (10X) form from Cell Signaling Technology. Follow these steps to ensure accurate and effective use of the buffer for your research needs.

Follow the steps to successfully fill out the RIPA Buffer (10X) form.

1. Press the ‘Get Form’ button to obtain the document and open it in the editing interface.
2. Read through the product description, which details that RIPA buffer is used for lysing cells and tissues. Ensure you understand the composition and recommended storage conditions.
3. Navigate to the section regarding the preparation of the buffer. Thaw the 10X buffer at 24-30°C while mixing end-over-end.
4. Dilute the 10X RIPA Buffer to a 1X solution by adding the appropriate volume of distilled deionized water (ddH2O). Note that this will yield enough 1X solution for making 150 mL of whole cell extract.
5. If using the buffer for adherent cells, wash the cells with phosphate-buffered saline (PBS) to remove any residual media before application.
6. Add 400 µL of the prepared 1X RIPA buffer to each 10 cm dish and incubate on ice for 5 minutes to allow proper lysis.
7. After incubation, scrape the cells from the plate to collect them.
8. Briefly sonicate the lysate to ensure thorough cell lysis.
9. Centrifuge the extract for 10 minutes at 14,000 x g in a cold microfuge to clarify the lysate.
10. Carefully remove the supernatant for further analysis or experimental application.
11. If necessary, repeat for non-adherent cells by adding 400 µL of buffer per 10^7 cells that have been washed and pelleted.
12. To store the buffer for future use, consider aliquoting the diluted solution and keeping it at -20°C.
13. Once all relevant steps have been completed, save changes, download the form, print it for your records, or share it as needed.

Get started now to complete your RIPA Buffer (10X) documentation online.