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CAS RN: | 產品號碼: N1208
Nuclear / Cytoplasmic Fractionation Kit
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補充產品訊息:
Product Information
- Components :1×Cytoplasmic Extraction Buffer, 20×Detergent solution, 1×Nuclear Extraction Buffer
- Size : 50 tests
Description
- This product is supplied for fractionating nuclear proteins and cytoplasmic proteins easily from cultured mammalian cells. Extracted protein can be used in downstream applications such as Western blotting.
Directions for Use
- [For Cytoplasmic Extraction]
- 1. Collect cells and wash twice with cold PBS. Use a cell scraper for adherent cells.
- 2. Resuspend the cells gently in cold 1×Cytoplasmic Extraction Buffer. Add 400 μL of the buffer to 100 μL of the cell volume (~ 1 × 107 cells).
- 3. Incubate on ice for 10 minutes.
- 4. Add 20×Detergent solution to the cell suspension and vortex vigorously for 10 seconds. Use 25 μL of the solution per 500 μL of the mixture.
- 5. Centrifuge at 800 × g for 10 minutes at 4 °C.
- 6. Transfer the supernatant (cytoplasmic fraction) to a new tube.
- 7. Resuspend the pellet in cold 1×Nuclear Extraction Buffer. Add 100 μL of the buffer to 100 μL of the pellet volume.
- 8. Incubate on ice for 20 minutes, vortexing every 5 minutes.
- 9. Centrifuge at 15,000 × g for 10 minutes at 4 °C.
- 10. Transfer the supernatant (nuclear fraction) to a new tube.
Storage
- Store at 4 °C.
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