An improved method combining two electrophoretic procedures: application to the separation of lens alpha-crystallin isoforms.
Polypeptides having different net electric charges and very similar molecular weights, visualized as one single band in sodium dodecyl sulfate--polyacrylamide gel electrophoresis (SDS-PAGE), can be readily analyzed by an improved method combining two electrophoretic procedures. The methodology consists of the identification and isolation of selected protein bands from SDS-PAGE, their equilibration in an isoelectric focusing (IEF) sample buffer, and their casting and separation in an IEF flat-bed gel. This method requires no extra equipment, is highly reproducible, is suitable for quantitative and comparative studies, and is especially useful in the case of small samples. As a particular example, we analyze here the subunit composition of alpha-crystallins of young and embryonic quail lenses.
Arruti
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Departamento de Histología y Embriología, Facultad de Medicina, Montevideo, Uruguay.
Chifflet
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D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D003459:Crystallins / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004625:Embryo, Nonmammalian; D007525:Isoelectric Focusing / Q000379:methods; D007908:Lens, Crystalline / Q000737:chemistry*; D011784:Quail
D003459:Crystallins