Quantification of Proteins on Western Blots Using ECL

  • Joanne Dickinson
  • Susan J. Fowler
Part of the Springer Protocols Handbooks book series (SPH)


The technique of Western blotting, the electrophoretic transfer of proteins from sodium dodecyl sulfate (SDS)-polyacrylamide gels to membranes, is a core technique in molecular and cell biology, see ref. 1 and  Chapters 39 42. Western blots are detected with antibodies specific to the target protein which are known as primary antibodies. A labeled secondary antibody is then added, which binds to the primary antibody. The label may be radioactive, colorimetric, fluorescent, or chemiluminescent. The major application of Western blotting has been the detection of individual proteins in complex mixtures such as cell lysates. Information can be obtained about the molecular weight of the protein of interest and its relative abundance in the sample. However, it is also possible to use Western blotting to provide accurate quantification of proteins in samples, to assess changes in protein expression levels (2).


Sodium Dodecyl Sulfate Amersham Pharmacia Biotech Accurate Quantification Charge Couple Device Camera Image Analysis Program 
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Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Joanne Dickinson
    • 1
  • Susan J. Fowler
    • 2
  1. 1.Amersham BiosciencesAmersham LabsUK
  2. 2.Amersham BiosciencesAmershamUK

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