Note

• This kit can extract the desired proteins from fresh or frozen plant fruits. However, some of the proteins may have reduced activity and may not be suitable for studies related to protein function.

Protocol

1. Grind 1-2 g of plant fruit sample in liquid nitrogen until it is pulverized.
2. Add 1 ml of Buffer A working solution and incubate at 65°C for 10 minutes, vortexing three times during incubation.
3. Centrifuge at 4°C, 20,000 rpm for 20 minutes and collect the supernatant.
4. Add 4 ml of Buffer B working solution to the supernatant and incubate at -20°C overnight.
5. Centrifuge at 4°C, 12,000 rpm for 10 minutes.
6. Add 10 µl of DTT (1M) and 10 µl of PMSF (0.1M) to each milliliter of cold Buffer C working solution, mix well, and keep on ice.
7. If the pellet is colored, repeat steps 4-5.
8. Add the Buffer C working solution kept on ice to the pellet to dissolve it.
9. Centrifuge at 4°C, 12,000 rpm for 10 minutes, and transfer the supernatant to a new tube.
10. Quantify the protein using the BCA method.
11. Aliquot and store at -70°C to avoid repeated freeze-thaw cycles.

Only for research and not intended for treatment of humans or animals

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