How to prepare a hemoglobin specimen for electrophoresis

Written by jess smith
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How to prepare a hemoglobin specimen for electrophoresis
Haemoglobin taken from blood samples can be analysed using gel electrophoresis. (Nicholas Eveleigh/Digital Vision/Getty Images)

Haemoglobin specimens can be obtained from an experimental subject or a patient and then separated into constituent parts using electrophoresis in a laboratory setting. Before haemoglobin can be run through gels using electrophoresis, it must be prepared so that it is properly and correctly separated during the process. This protocol assumes that the correct gels and appropriate electrophoresis techniques will be used, according to the desired analysis.

Skill level:
Moderately Challenging

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Things you need

  • Haemoglobin sample (from blood)
  • Eppendorf tubes
  • Laboratory freezer at --6.67 degrees C centrigrade
  • Gel Electrophoresis kit

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Instructions

    Preparation

  1. 1

    Transfer the whole blood sample to be used for electrophoresis into an eppendorf tube. Ensure the tube is properly closed.

  2. 2

    Freeze this sample in a freezer suitable for the storage of organic materials such as blood, at --6.67 degrees C centrigrade. Store until completely frozen (a few hours or perhaps overnight).

  3. 3

    Defrost the sample, without vortexing or mixing, and remove the supernatant; this is the hemolysate that will be used for electrophoresis.

  4. 4

    Add the buffers most suited to the gel and electrophoresis conditions to be to run the supernatant sample.

  5. 5

    Load and run the gel electrophoresis experiment according to the equipment and gel being used.

Tips and warnings

  • This method can also be used if the blood sample is in the form of blood clots or separated red blood cells.
  • Blood samples must only be handled by qualified and trained persons, in a clinical or scientific setting.
  • Ensure that appropriate precautions are taken when working with blood samples.

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