Hi everyone,
Currently, I am doing ChIP assay using EpiQuik Plant ChIP kit from Epigentek. At step "Antibody Binding to the Assay plate", the kit instruction says that the strip wells must be washed by CP1 buffer. However, I have never used this kit previously so I do not know how to do this washing step well. Should I just add the CP1 buffer into the well and pipetting several times [how many time? I do not know either], and then withdraw the CP1 buffer by using a pipette?
And I am also wondering whether this kit works better than other manual protocols such as using salmon sperm DNA/protein A agarose beads to catch the immunoprecipitated complex.
I am really wondering to hear your answers and thank you guys very much! :)
I don't know the kit, but washing wells on a 96 well plate (I guess it's 96 well plate format?) normally consists of pipetting the wash buffer into the well and immediately removing it afterwards. If the protocol does not further precise the step, this procedure should be fine.
I'm also interested in the efficiency of the kit, since I'm normally using Protein G covered agarose or magnetic beads for ChIP.
Hi there! Thanks for using our kit.
If you have not started or finished the kit procedure yet, to answer your question, it is recommended to pipette the wash buffer into the well and immediately aspirating it out, so Ariane is correct! Although this is for a different kit, you can see the wash step demonstrated in our MethylFlash kit video at the 1:24 mark: https://www.youtube.com/watch?v=7uI1k3DzFuA
Good luck -- and I hope you'll let us know how the kit performed.
PS Ariane, if you are further interested in kit information, feel free to reach out to us!
Hi Nguyen,
Thank you so much for the information. I am glad to hear that the kit worked well -- we recognize the concern about the need for extra buffers as other customers have brought this to our attention before. In the event we release the next or updated version of a plant ChIP kit, we will certainly factor in the reagent amount requirements.
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