I have been attempting to extract a protein expressed using IPTG induction from my BL21 culture through sonication for an extended period, but this method has repeatedly failed. Subsequently, I tried periplasmic extraction by inducing osmotic shock using sucrose, which also did not result in the release of the protein. Despite troubleshooting the extraction with various protocols, the problem persists. I would greatly appreciate any advice on the procedure or alternative approaches to address this issue. Below are the details of sonication i used for my protocol :
- Sonication: 30% / 40% amplitude with 30-second pulses ON, followed by a 30-second cooling period, repeated for 8-10 cycles.
- Also, the plasmid used for the expression uses signal peptide pelB with binding modules for cellulose.
I've attached images showing the sonication results (pellet and supernatant with minimal protein) and the periplasmic extraction.
- Badges
- Science topic
- Similar topics
- Mathematical Sciences
- Graphs
More Vaibhav Tyagi's questions See All
Similar questions and discussions