Bacterial Protein Extraction and Purification
Corresponding Organization : University of Chieti-Pescara
Variable analysis
- Lysis using a Q Proteome Bacterial Protein Prep kit containing 100 mg/mL lysozyme
- Addition of Benzonase 0.25 U/µL to degrade nucleic acids
- Protein concentration of samples (8 µg/µL for planktonic and 16 µg/µL for biofilm)
- Incubation at room temperature on ice
- Centrifugation at 14,000× g for 30 min at 4 °C
- Centrifugation at 50,000× g for 30 min
- SpeedVac drying and storage at -80 °C until protein purification
- Pooling of three independent extractions for planktonic and biofilm samples
- Protein purification using a Ready Prep 2D Cleanup kit
- Protein resuspension in 2-DE solubilization buffer
- Centrifugation at 12,000× g
Annotations
Based on most similar protocols
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!