This kit provides a full set of reagents, suitable for preparing cytoplasmic proteins from cells and animal tissues. The extraction process is simple and convenient and can be completed within 1 hour.The prepared cytoplasmic protein not only has high purity and maintains natural activity, but also rarely cross-contaminates. The extracted protein can be used for Western Blotting, transcriptional activity analysisProtein studies such as analysis, gel shift gel block experiment, immunoprecipitation, and enzyme activity determination.

Instructions: Precautions for use:

1. The reagents in the spin cap centrifuge tube should be centrifuged briefly before opening the cap, and the liquid on the inner wall of the cap should be thrown to the bottom of the tube to avoid liquid spillage when opening the cap.

2. All reagents during the experiment must be pre-cooled; all appliances must be pre-cooled in a -20℃ refrigerator. The sample must be kept at low temperature throughout the process. A. Cellular protein extraction 1. Take 5-10×106 cells, centrifuge at 500×g at 4°C for 2-3 minutes, carefully aspirate the culture medium, as much as possible, and collect the cells. 2. Wash the cells twice with cold PBS. After each wash, absorb the supernatant as much as possible.

3. Add 200 μl of cold cytoplasmic protein extract, 20 μl of protease inhibitor mixture to every 20 ul cell volume, vortex and vortex at high speed for 15 seconds,Place on ice for 10 minutes.

4. Vortex again at high speed for 5 seconds, and then centrifuge at 16000×g at 4°C for 5 minutes.

5. Quickly draw the supernatant into another pre-cooled clean centrifuge tube to obtain cytoplasmic protein. Please store on ice or in a refrigerator at -80℃ for future use. B. Tissue protein extraction 1. Take an appropriate amount of tissue sample and shred it, add cold PBS, homogenize with a tissue homogenizer until there is no visible solid (or grind with liquid nitrogen), static on ice Incubate for 5 minutes and carefully draw the supernatant into another pre-cooled clean centrifuge tube. 2. Centrifuge at 500×g at 4°C for 2-3 minutes, and discard the supernatant. 3. Follow the third step of the cell protein extraction method. Please store the above protein extract in a refrigerator at -80℃ for later use