I am planning to perform a co-immunoprecipitation experiment using the Pierce His Protein Pull-Down Kit (https://www.thermofisher.com/order/catalog/product/21277). My bait and prey proteins are both previously purified, and my bait protein as an N-terminal His tag. I am trying to determine what the appropriate controls would be to ensure that any interaction I see between and bait and prey is specific. The kit mentions that I should do a "Minus bait, Plus prey" control, which would determine if the prey interacts non-specifically with the beads. They also mentioned a "Plus bait, Minus Prey" control. Are there any other controls I should plan for?
It's great that you're planning ahead and considering controls before you are starting to collect data.
Read through their supporting materials: https://www.thermofisher.com/us/en/home/technical-resources/technical-reference-library/protein-purification-isolation-support-center/protein-ip-coip-pulldown-support.html
And find a few publications that use co-ip and see what controls they used, it will depend a bit on the particular experiment.
Good luck!
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Materials Science
- Materials
- Inorganic Materials
- Metals
- Alloys
- Steel