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Hi,
I am working for BDNF measurement in human bloods.For this, I need to get my capture and detection Ab's working. My detection Ab is mouse monoclonal biotin conjugated. One-site ELISA worked for my biotin conjugated Mab. For last couple of weeks I have been optimizing the concentration of Mab's. My concentration for capture is 3ug/ml and for detection Ab is 2ug/ml. My problem is that, at lowest concentration of detection Ab, my blank is too high. I tried using different blocking buffers, washing buffer. My supervisor says that I dont have any problems with buffer. Does Acetic Acid interfere the biotin? Can you guys please help me out with the biotin conjugated detection Ab (mouse monoclonal) protocol?
Thank you.
