It doesn’t matter which chemiluminescent substrate you use for your Western, right?
Wrong!
Maybe back in the early days of chemiluminescent Westerns one substrate was all there was. But these days different chemiluminescent substrates are designed for different purposes.
For example, let’s take a look at Advansta’s 3 WesternBright chemiluminescent substrates.
WesternBright ECL
Looking for an all-purpose substrate for routine detection of proteins? Then WesternBright ECL can do the job for you.
ECL is a sensitive substrate perfect for general film imaging. Using this substrate you can detect low pg amounts of protein while using up to 10x less antibody. The long-lasting signal lets you take your time to get the perfect exposure.
ECL even comes as a convenient spray! Just point the bottle containing the premixed solution at your blot and spray.
WesternBright Quantum
Do you have quantitation on your mind? If you do, you might want to give WesternBright Quantum a try.
Quantum was specifically designed to give the broadest useful linear range when using CCD imagers. Quantum has a high sensitivity for low abundance proteins, but doesn’t exhibit substrate depletion when detecting high abundance proteins. This leads to a linear range over 3 orders of magnitude!
Quantum’s long-lasting signal (you can image hours after the addition of substrate) was optimized for CCD imagers, but can also be used with film.
WesternBright Sirius
WesternBright Sirius is a seriously sensitive substrate.
Sirius was designed for the difficult-to-detect protein. With a high signal-to-noise ratio and low background, Sirius is Advansta’s most sensitive substrate. It is ideal for detecting low abundance proteins or when using a very low concentration of antibody.
Sirius can be used with both film and CCD imagers.
Of course, you should always do the actual experiment to determine which substrate works best with your protein, but as a general rule, you can remember:
- Need an all-purpose substrate for film imaging? – Excel with ECL
- Quantitation on your mind? – Give Quantum a try
- Need serious sensitivity? – Stick with Sirius
Photo courtesy of Incase.
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