Antibodies 101: The Basics of Western Blotting
Type
Blog Post
...commonly used, PVDF tends to be slightly more popular due to its durability and high binding capacity. In addition... (HRP), that emits light upon reacting with a specific substrate. The emitted light, or chemiluminescence... samples, a protein standard of known molecular weight in a separate lane can help confirm the size of...acrylamide content of the gel. For small proteins, use a higher percentage of acrylamide to increase migration ...broad range of protein sizes but takes hours to overnight.
In contrast, a semi-dry transfer only requires...chain. Consequently, antibodies validated for IP might not work for a western and vice versa.
In addition...specific experimental condition you could look for a slight size shift in the protein band before and after...