A novel method that improves sensitivity of protein detection in PAGE and Western blot

作者：Vallejo Illarramendi Ainara^*; Marciano Denise K; Reichardt Louis F

来源：Electrophoresis, 2013, 34(8): 1148-1150.

DOI：10.1002/elps.201200534

• Microloader
• Phosphoproteins
• Polyacrylamide gels
• Western blot

We have developed a simple and inexpensive method that improves sensitivity of protein and antigen detection in standard PAGE procedures. Our technique uses a sample microloader device with a funnel-like structure, filled with a 4% stacking gel. When attach to the top of a polyacrylamide slab gel, the proteins in a sample are concentrated by electrophoresis into a small volume as they emerge from the device's narrow outlet. Our microloader has several advantages over previous devices, including simple assembly, high versatility, and absence of cross-contamination between lanes. Addition of this device to a slab gel results in a fivefold increase in the sensitivity of antigen detection in a Western blot. As a result, less protein is required for loading and signal detection. Our protocol is a straightforward modification of a standard experimental technique, and is especially useful when only limited sample quantities are available.

• 出版日期2013-4