The Protein Man's Blog | A Discussion of Protein Research

There are a lot of reasons why researchers all over the world choose enhanced chemiluminescence (ECL) detection for a wide range of Western blot applications. ECL is considered as the experts’ detection method of choice due to its high sensitivity, excellent signal-to-noise ratio, and wide dynamic range. Additionally, ECL is also useful in quantifying a wide variety of biological materials such as cell RNA, DNA and other analytes. 

After running your gel and transferring your proteins to your membrane, there is one thing left for you to do to ensure the accurate analysis of your protein. You need to block all unoccupied sites on your membrane to prevent the non-specific binding of antibodies and other detection agents to your membrane during subsequent steps. If you take this step lightly, you'll risk compromising the reliability your results.

Western blotting is an immunodetection technique used to first separate proteins by gel electrophoresis then to transfer those separated proteins from the gel matrix onto a secondary, more durable matrix, most commonly polyvinylidene difluoride (PVDF) or nitrocellulose.

The choice of constant current or constant voltage will depend on several different factors that must be taken into consideration for each Western transfer test. The level of heat, duration of the test, type of proteins, the size of the transfer, and the rate of transfer will all be affected by power, voltage, and current being used to drive the test. Each factor will play a role in how well the proteins transfer and the ultimate accuracy of the test.