Electrophoretic separation of LDH isoenzymes on cellulose acetate.
Virtual experiment.

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m1 m2 m3
A
B

a new electrophoresis (with the same sample tubes)

Fundamentals: On this page you can work to understand the principles of the analyisis of lactate dehydrogenase (LDH) isoenzymes using electrophoresis.

Samples m1, m2 and m3 for this experiment are the supernatants obtained after homogenisation of tissues which contain lactate dehydrogenase and centrifugation to remove any insoluble material.

What are Pasteur pipettes and how they are used (video)

Instructions

  1. Drag the pipette to one of the tubes containing a sample.
  2. Drag the pipette positioning it on the centre of one of the cellulose acetate strips situated on the electrophoresis tank, so that the sample is applied.
  3. Drag the pipette to the waste container (a new pipette will appear later on the left)
  4. Repeat this process with a new sample, applying it on the other acetate strip.
  5. Press the switch in the power source.

In your lab notebook: wrie down the code assigned to your experiment (in this case it is and which sample (m1, m2 or m3) you have applied to each strip (A and B). Then, copy the images with the results (stained strip and densitogram). Finally, interpret results: from which tissue has each sample been prepared?

Results after running the electrophoresis and applying the satining which detects LDH enzyme activiy:

PMS = phenazine methosulphate
NBT = nitro blue tetrazolium
Versión 2.0
(+)
(–)
Densitogram:
Cellulose acetate strip:
 
captured images:
Author: Angel Herráez. Part of the Biomodel.uah.es website

CC by-nc-sa Offered to be used uner the terms of the Creative Commons Attribution – NonComercial – ShareAlike License.

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