Technical Design of a Western Blot
Type
Blog Post
...assay run time to two days.
Indirect or direct detection method
Indirect detection is the most common...quicker to use, with high binding capacity, though it has lower retention after binding and washes. It’s less...for validation data, or follow the references to find the original publication and ensure that it includes...for a western blot. Polyclonal antibodies, which bind to multiple sites on an antigen, are cheap and can...the manufacturer, each rAb in a mixture can be individually validated. rAbs are not as commonly available...). Direct westerns are simpler and faster than indirect ones and can be used in highly quantitative assays...tested to ensure it doesn’t impact the antibody’s binding to your protein of interest.
Protein preparation...