title: A novel method that improves sensitivity of protein detection in PAGE and Western blot.
abstract: 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.
authors: Department of Physiology, University of California, San Francisco, CA, USA. ainaravallejo@yahoo.es
authors: Ainara
authors: A
authors: Vallejo-Illarramendi
authors: Denise K
authors: DK
authors: Marciano
authors: Louis F
authors: LF
authors: Reichardt
mesh_terms: D015153:Blotting, Western / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation* / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D012680:Sensitivity and Specificity
chemical_list: D011506:Proteins
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