Hi all,
I am trying to detect biotinylated DNA that has been transferred onto nylon membranes by chemiluminescent with biotin antibodies. However, I am getting very high background in my results, which I believe is due to inadequate blocking. I understand that milk is not a suitable blocking agent as it contains biotin so I tried with 5% BSA to no avail. Is there any other blocking agent that will be effective? Thanks in advance!
In my research, I use a antibody against 5-methyl-cytosine. To block our membrane before probing with the antibody, I block the membrane with 10% low fat milk during one night at 4°C. I have no background.
You could try gelatin or BSA and increase the time of blocking to one night.
Dear Xi Xiao Huang , I am wondering if you have figured out how to block your membrane? I am having the same issue and any suggestions will be appreciated.
Thank you!
Could you share your protocol to detect biotinylated DNA that has been transferred onto nylon membranes by chemiluminescent with antibodies?
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