title: Comparison of Stain-Free gels with traditional immunoblot loading control methodology.
abstract: Loading controls are necessary for semiquantitative Western blotting to compensate for loading errors. Loading control methods include the reprobing of membranes with an antibody against a constitutively expressed protein or staining the membrane with a total protein stain. We compared the loading control performance of recently released Stain-Free (SF) gels with Sypro Ruby (SR) and reprobing using β-actin. SF gels demonstrated superior performance in that they were faster, required fewer steps and consumables, and allowed the quality of electrophoresis and Western transfer to be assessed before committing to costly and time-consuming Western blots.
authors.affiliation: Department of Ophthalmology, Flinders University, Bedford Park, SA 5042, Australia. alex.colella@flinders.edu.au
authors.forename: Alex D
authors.initials: AD
authors.lastname: Colella
authors.forename: Nusha
authors.initials: N
authors.lastname: Chegenii
authors.forename: Melinda N
authors.initials: MN
authors.lastname: Tea
authors.forename: Ian L
authors.initials: IL
authors.lastname: Gibbins
authors.forename: Keryn A
authors.initials: KA
authors.lastname: Williams
authors.forename: Tim K
authors.initials: TK
authors.lastname: Chataway
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D000906:Antibodies / Q000276:immunology; D015153:Blotting, Western; D004396:Coloring Agents / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting / Q000295:instrumentation; D009942:Organometallic Compounds / Q000737:chemistry; D051381:Rats; D012160:Retina / Q000378:metabolism; D059629:Signal-To-Noise Ratio
chemical_list: D000199:Actins; D000906:Antibodies; D004396:Coloring Agents; D009942:Organometallic Compounds; C515601:Sypro Ruby