Hello everyone,
I was doing dot blot technique for detection of Iba1 in my sample. The first time, the image background wasn't very helpful for further analysis. So, I continued with b-actin detection (where the images were perfect) and then I repeated with the first antibodies (Iba1 detection) where the images were perfect as well. I have to mention that all my antibodies have antimouse HRP antibody as a secondary one. As a result, I am anxious whether the second image of Iba1 detection is real or not. The stripping protocole I use is the mild one seen below. What do you think? Should I change my stripping protocole or just detect b-actin as a last step? Any recommendations are welcome.
Thank you very much in advance.
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