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In this section, you can find solutions to issues related to protein band size and pattern problems. Explore possible causes and solutions for bands appearing very high or very low in western blot. Check the expiration date on the protein standard
Expired lots may result in faded or missing bands due to protein degradation Explore our troubleshooting tips from experts on western blot issues like no signal, high background, multiple bands, and more Check the storage conditions for the protein standard
Improper storage conditions will compromise the stability of the proteins in the standard.
If all the bands on your blot, including the molecular weight ladder, are difficult to see, it could indicate a problem with your technique rather than the protein you're trying to detect Familiarize yourself with the western blot protocol and check the common pitfalls below. In this western blot troubleshooting section, we will help you visually identify specific and common problems on your western blots, such as high background, weak or no signal, multiple bands, uneven staining and suggest what may be causing them and some solutions to remedy them. When none of the protein bands appear on the membrane, the most likely reason is problems relating to either the equipment or the assembly of the gel membrane sandwich
The polyvinylidene fluoride (pvdf) or nitrocellulose (nc) membrane should always be oriented on the anode (+) side of the gel. Variability between antibody batches or antibody degradation due to expiration can result in inconsistent detection. Problems, causes and solutions for issues in western blot
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