Electrophoresis of a-amylase and invertase Creative Commons License This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. This note on methods is available in Fungal Genetics Reports: http://newprairiepress.org/fgr/vol9/iss1/2 Grotzner, H. G. A convenient method for a-amylose Cellulose acetate serves as an attmctive electrophoretic medium for o-rid invertare electrophoresir on cellulose cc&ate strips. mpid separation of enzyme proteins. However, difficulty is

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... tered when one wishes to vitalize an enzwne on the rtrio. For this reason, we have developed a simple technique for we with a-omylase from Neuroyrom culture media. The culture medium (Vogel's minimal, or phosphate buffer) from induction experimenh is placed into a dialyxir sack and concentrated qoinst solid sucrose in the cold room overnight (Horowitz and Fling I962 NeumrpDro Newrl. 2: 19). Twelve lambda samples of this concentrate are placed on pre-sooked, blotted reprophore 111 (Gelman) strips at D position new the cathode and electmphorized for I how ot 300 v (or approximately 75 volts/strip). The electrolyte is 0.3 M borate buffer, pH 8.2. At termination of the run, the strips ore blotted and placed upon 0.5% starch -2% agor blocb and incubated for about IO minutes. The strips ore then peeled from the ogor blocks and stained in iodine vapor for several minutes. This procedure giver o smooth, blue background with white bands corresponding to omylase activity. The block. con also be stained as a duplicate. For the simultaneous localization of invertare, the strip, or o longitudinal holf of the amylore strip, is sliced into 3 mm lateml strips, which are then sequentially eluted in small tuber with distilled water. An equal volume of buffered, IO-2 M sucrose is odded to the tuber, and the tubes are incubated at 3PC for I hour. Reducing sugar is assayed in each tube by means of o-nitroralicylate reagent (Bernfeld, in Methods in Enzymology, Vol. I).