Plant protein extraction method summary
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July 10 13:32:27, 2025
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**Summary of Plant Protein Extraction Method**
This method is commonly used for extracting proteins from plant tissues, especially when preparing samples for SDS-PAGE or vertical gel electrophoresis. The procedure involves several key steps that ensure the efficient and reliable extraction of high-quality protein.
First, based on the sample weight (typically 1 g of tissue is mixed with 3.5 ml of extraction buffer), prepare the extraction solution on ice. The exact volume can be adjusted depending on the type of plant material being used. It's important to keep the buffer cold throughout the process to prevent protein degradation.
Next, place the plant tissue in a mortar and grind it thoroughly using liquid nitrogen. This helps to break down the cell walls and release the proteins effectively. Once the tissue is ground into a fine powder, transfer it into the pre-chilled extraction buffer and let it stand on ice for 3–4 hours. This incubation allows the proteins to be solubilized and released into the buffer.
Afterward, centrifuge the mixture at either 8000 rpm for 40 minutes at 4°C or 11100 rpm for 20 minutes at 4°C, depending on the centrifuge available. The supernatant obtained after centrifugation contains the extracted proteins and can be collected for further analysis.
The extraction buffer composition is as follows:
- 300 ml of 1 M Tris-HCl (pH 8.0)
- 45 ml of glycerol
- 75 ml of polyvinylpyrrolidone (PVP)
- 6 g of PVP (optional, depending on the plant species)
This protocol is particularly suitable for SDS-PAGE and vertical slab gel electrophoresis. It ensures good resolution and clarity of protein bands, making it ideal for downstream applications such as Western blotting or mass spectrometry.
Make sure all reagents are prepared fresh and stored properly to maintain their effectiveness. Always work under sterile conditions and avoid contamination during the process.
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