National Centre for Biotechnology Education Bacterial Transformation Copyright © Dean Madden, 2013.
National Centre for Biotechnology Education Gel electrophoresis Protein power! Copyright © Dean...
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National Centre for Biotechnology Education www.ncbe.reading.ac.uk Gel electrophoresis Protein power! Copyright © Dean Madden, 2010
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Transcript of National Centre for Biotechnology Education Gel electrophoresis Protein power! Copyright © Dean...
National Centre for Biotechnology Education
www.ncbe.reading.ac.uk
Gel electrophoresis
Protein power!
Copyright © Dean Madden, 2010
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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
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Fit a tip firmly onto a syringe, using some tubing as an adapter
Use this syringe and tip to transfer
your protein samples
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Add 0.5 mL of blue marker dye to each protein sample
About 0.5 mL
Protein sample
Label the tube
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Float the tubes in boiling water for three minutes, then store them on ice
OPTIONAL STEP
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Frosted panel on this side
Molten agarose (3%)55–60 °C
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Cut two electrodes
Carbon fibretissue
42 mm
22 mm
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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
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Electrodes
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Direction of protein movement
Place a comb over the tank to reduce
evaporation
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Leave the stain on for 60 minutes
Coomassie blue stain
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The Coomassie blue stain soaks into the gel and
reveals the proteins
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Wells Marker dye
Protein bands
