Due to its high sensitivity, wide dynamic range, and high signal-to-noise ratio, enhanced chemiluminescence or ECL is considered as one of the most popular detection methods for a variety of western blotting applications in most protein laboratories around the world. This method also proves to be very useful in the quantification of a wide variety of biological materials such as cells, proteins, RNA, DNA, and a host of other analytes.
Topics: Western Blotting
INTRODUCTION:
PVDF and nitrocellulose membranes are both used in Western blotting and have various different characteristics, however a common question asked is "Why do PVDF membranes require a methanol soak?".
Check out this blog for more on the differences of the membranes:
Topics: Western Blotting
When doing a Western blot procedure, the macromolecules are first separated using gel electrophoresis. The separated macromolecules are transferred onto a second matrix (nitrocellulose or polyvinylidene difluoride (PVDF) membrane) and the membrane is blocked to prevent the nonspecific binding of antibodies to its surface. The transferred protein is then complexed with an enzyme-labeled antibody (probe) and an appropriate substrate is added to produce a detectable product.
Topics: Western Blotting, Protein Detection
When it comes to Western blotting, choosing the right membrane for your application usually spells the difference between success and failure. However, this is usually easier said than done since you need to consider the properties of your protein and the downstream detection steps required in your application before you can determine which membrane can give you the results that you need.
Topics: Western Blotting
