Notes

• This kit can extract the desired proteins from fresh, frozen, or dried plant tissues, including leaves, roots, flowers, and seeds. Most of the extracted proteins maintain their activity. However, proteins extracted from dried plants or tissues ground in liquid nitrogen may have reduced activity and may not be suitable for studies related to protein function.

• To prevent protein precipitation or denaturation, dialysis, solution exchange, or the addition of a protein stabilizer is recommended. For short-term storage, add PEG or glycerol to a final concentration of 5-10%. For long-term storage, add PEG or glycerol to a final concentration of 10-20%.

Protocol

Lysis Buffer Working Solution Preparation:

Prepare the working solution by adding 10 µl of protease inhibitor, 10 µl of DTT (1M), and 10 µl of PMSF (100 mM) to each milliliter of cold Lysis Buffer. Mix well and keep on ice until use.

Procedure:

1. Grind 100-200 mg of plant sample in liquid nitrogen until it is pulverized.
2. Add 1 ml of the Lysis Buffer working solution to the ground sample. Incubate at 4°C for 30 minutes, vortexing three times during the incubation.
3. Centrifuge at 4°C, 10,000 rpm for 20 minutes. Collect the supernatant as the protein extract and quantify using the BCA protein assay.
4. 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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