National Centre for Biotechnology Education www.ncbe.reading.ac.uk Gel electrophoresis Protein power! Copyright © Dean Madden, 2010.

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Presentation transcript:

National Centre for Biotechnology Education Gel electrophoresis Protein power! Copyright © Dean Madden, 2010

The negatively-charged proteins move through the gel towards the positive electrode The gel contain minute holes, so it acts like a sieve. Small proteins move quickly; larger proteins follow on behind. Positive electrodeNegative electrode

Copyright © Dean Madden, 2010 Fit a tip firmly onto a syringe, using some tubing as an adapter Use this syringe and tip to transfer your protein samples

Copyright © Dean Madden, 2010 Add 0.5 mL of blue marker dye to each protein sample About 0.5 mL Protein sample Label the tube

Copyright © Dean Madden, 2010 Float the tubes in boiling water for three minutes, then store them on ice OPTIONAL STEP

Copyright © Dean Madden, 2010 Frosted panel on this side Molten agarose (3%) 55–60 °C

Copyright © Dean Madden, 2010 Cut two electrodes Carbon fibre tissue 42 mm 22 mm

Copyright © Dean Madden, 2010

Use a fresh tip for each sample you load into the gel Label the end of the tank to show the contents of each well Black card under the tank reveals the wells for loading 2–3 mm depth of buffer over the gel

Copyright © Dean Madden, 2010 Electrodes

Copyright © Dean Madden, 2010 Direction of protein movement Place a comb over the tank to reduce evaporation

Copyright © Dean Madden, 2010 Leave the stain on for 60 minutes Coomassie blue stain

Copyright © Dean Madden, 2010 The Coomassie blue stain soaks into the gel and reveals the proteins

Copyright © Dean Madden, 2010 WellsMarker dye Protein bands