Dear all,
I am looking for a commercially available pull down assay kit that has conducive buffer conditions and high affinity for the antibody conjugation. My protein of interest (bait) shows a very low binding with the prey and so is there any enhancer that can give good signal for sub-nanomolar concentration ? As I have been using protein A dyna bead from Invitrogen, and every time I see non specific heavy chain of the protein Ig in the western blot. Then to get around the later issue, I used True Blot secondary Antibody, however I can still see huge blob for the Ig heavy chain from the bead. So, if any one can suggest to sort out these two issues with my pull down assay, it would be a great help.
Thank you
With kind regards
Prem
Dear Prem Prakash
You can use Chromotek GFP-trap magnetic agarose beads for the pull-down/co-immunoprecipitation assays. These magnetic beads are coupled to GFP nanobody and are ready to use. Only you need to label your bait protein with GFP. This product has recommended set of buffers but you can modify them according to your model organism and condition.
Best wishes.
You could also think about using PICO to detect your protein of interest.
Here is a video how it works and the link to the shop:
https://www.youtube.com/watch?v=ptZx-IgLW3I
Marius Pollet Thank you so very much it seems very promising ! Could you please suggest any research article that describes the principles, methods and results of PICO. That would be a great help.
Thank you
With kind regards
Prem
You can find it here: figure 2 j
https://www.nature.com/articles/s41586-019-0955-3
Since Actome cooperates with qiagen its called PICO in the paper it's called "emulsion coupling"
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts