使用菌株培养菌体后如何进行蛋白质组分析?
Answer:
- Harvest and lyse the bacterial cells.
- Centrifuge the cell lysate.
- Collect the pellet and resuspend it in a lysis buffer.
- Add protein extraction reagents to the lysate.
- Incubate the lysate at 56°C for 1 hour.
- Centrifuge the lysate at 12,000 × g for 10 minutes.
- Collect the supernatant.
- Dry the supernatant and resuspend it in a protein sample buffer.
- Run the protein sample on an SDS-PAGE gel.
- Analyze the protein bands on the gel.
Additional notes:
- The specific protein extraction reagents to be used will depend on the protein targets of interest.
- The incubation time and temperature may also need to be optimized for each protein.
- The protein bands on the SDS-PAGE gel can be analyzed using a variety of methods, such as Western blotting, mass spectrometry, or immunoprecipitation.