255 - Frederiksen

Purification & Size Determination of gfp & bfpEDVO-Kit #25515Staining the Gel - OPTION #2Staining with Protein Plus stain (liquid)1. Use a small plastic tray or large weigh boat for staining the gel individually.2. Remove the gel cassette from the electrophoresis apparatus and blot offexcess buffer with a paper towel.3. Lay the cassette down and remove the front plate by placing a spatulaor finger at the top edge, near the sample wells, and lifting it away fromthe larger back plate. In most cases, the gel will stay on the back plate.If it partially pulls away with the front plate, let it fall onto the backplate.4. Slide the gel into a staining tray.If the gel sticks to the plate, pipet some of prepared staining solutiononto the gel and gently nudge the gel off the plate.5. Cover the gel with 50ml of prepared staining solution. The gel shouldbe submerged in stain.The Experiment6. Cover the staining tray with plastic wrap to prevent evaporation. Stainfor approximately two hours. The gel can be stained in a few hoursor overnight but will require a longer destain. (Note: The gel willshrink.)Destaining the gel7. Pour off the stain from the gel in the staining tray.8. Add 100ml of destaining solution. The gel should be submerged underdestaining solution.Gentle agitation or incubationon a slow rotating orshaking platform will acceleratethe destain time.9. Destain overnight (12 to 14 hours).10. Remove the gel from the destaining liquid.11. View the gel on a white light source. Photography of protein bands isoptional.12. Examine the protein bands from the various samples and approximatetheir molecular weights by comparison with the standard protein markers.Storing the GelThe gel should be soaked in a mixture of 50ml of distilled water containing6ml of acetic acid and 3ml of glycerol overnight. The gel can be stored inthis solution or dried as described on page 13.Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratoryuse only. This document, or any part, may not be reproduced or distributed for any other purpose without thewritten consent of EDVOTEK, Inc. Copyright © 1998, 2000, 2001, 2005, 2007 EDVOTEK, Inc., all rights reservedEVT 005097KThe Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com