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FAQs
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What is a chromogenic substrate?
Chromogenic substrates used in western blotting allow for direct visualization of a target protein. They work by reacting with an enzyme such as HRP or AP attached to the detecting antibody, producing an insoluble, colored precipitate on the membrane.
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When should I choose HRP vs AP?
Choose HRP for a fast, cost-effective, general-purpose method for presence/absence of protein. Select Alkaline phosphatase (AP) when stability of the signal is a priority or when multiplexing different substrates.
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When should I choose chemiluminescent vs. colorimetric method for western blotting?
Choose the chemiluminescent method when you need to detect low-abundance proteins because it offers high sensitivity, capable of detecting signals in the femtogram range. Choose the colorimetric method when you have high-abundance proteins and prefer a faster, simpler, and less expensive method that doesn't require specialized imaging equipment.
