Hello Everyone!
Recently i tried Mito protein western blot from whole lysate of mice tissue for one of the subunit of COX 8A (Size= 8KD) and One subunit of NADH dehydrogenase (SIZE = 17KD) NDUFA12.
I am following all the steps correctly, but bands are only showing around 10 KD in case of both the antibodies (Gels run separately for both the antibodies).
I am wondering, if somebody have used the same antibodies.
Thanks in advance
There are a few potential explanations for why you might be seeing bands at a lower molecular weight than expected on your Western blot. Some possibilities include:
To troubleshoot the issue, it can be helpful to repeat the Western blot with fresh samples and reagents, and to include positive and negative control samples to ensure that the assay is working correctly. Additionally, checking the specificity of the antibodies you are using and optimizing the sample preparation process can help to improve the reliability of your results.
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