I want to lyse bacteria using beads for ELISA. Campylobacter to be specific. How long do I have to lyse it for and at what speed and time of centrifugation after?
Dear Theros,
I suggest 4,500 rpm, 6 x 20 sec with 1 min on-ice intervals. Thereafter, Centrifuge at 8,000 g at 4 °C for 5 min.
Good luck.
Hello, Theros,
It depends on the bacterial strains, the size of the beads, temperature of the shaking, and the protein stability of what you have in mind. I would suggest to set up a small experiment based on the published data to examine different shaking conditions, and centrifugation conditions for your use. After collected the lysates, the most suitable condition will be decided on the total protein in the lysate and the levels of protein of choice.
Good Luck
We use sonication to produce lysates of Salmonella and Campylobacter for use in ELISA as antigen. We do a fair bit of downstream processing to produce soluble lysates that can be used in both ELISA and cell-based assays. It is a little laborious but you can make enough in batch to last a couple of years.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA