Horizontal gel electrophoresis with sample volumes up to 1.5 mL, using a discontinuous buffer system and automated apparatus.
Horizontal gel electrophoresis has previously suffered from lack of an instrumental design that would allow for application of large sample volumes. A recently introduced commercial apparatus (HPGE-1000, LabIntelligence) remedies that problem in application to gel electrophoresis with intermittent scanning of fluorescence by using the concentrator module of that apparatus as a stacking gel reservoir. By using this technique and a 3 mL stacking gel, protein samples of up to 1.5 mL yield bands in the horizontal resolving gel that are independent of sample volume in their width, area and migration rate. A remaining procedural problem relates to the apparent nonsimultaneity of dye and protein entrance into the resolving gel, which necessitates peak characterization by absolute rather than relative mobilities.
D001331:Automation; D002021:Buffers; D003207:Conalbumin / Q000032:analysis*; D004589:Electrophoresis, Disc / Q000379:methods*; D005449:Fluorenes; D005456:Fluorescent Dyes; D007477:Ions; D013388:Succinimides
D002021:Buffers; D005449:Fluorenes; D005456:Fluorescent Dyes; D007477:Ions; D013388:Succinimides; D003207:Conalbumin