Autoradiography of 2-D Gels
By: Andrew J. Link2
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Autoradiography is used to visualize and quantitate radiolabeled proteins that are resolved by 2-D protein gel electrophoresis.
Proteins are commonly labeled in vivo with either 3H, 14C, 35S (low-energy (β-emitters), 32P (high-energy β-emitter), or 125I (high-energy γ-rays). During film-based autoradiography, these emitted particles or γ-rays enter the film and cause the
ejection of electrons from silver halide crystals generating local precipitates of silver atoms. In the past several years,
the use of phosphor imaging has been replacing film-based autoradiography. Phosphor imaging has 10–250 times increased sensitivity
compared to film-base autoradiography. While film has a dynamic range of about 300 to 1, phosphor imaging has a linear dynamic
range over several orders of magnitude (1).
Book Title: 2-D Proteome Analysis Protocols
Series: Methods in Molecular Biology | Volume: 112 | Pub. Date: Sep-24-1998 | Page Range: 285-290 | DOI: 10.1385/1-59259-584-7:285
Subject: Biochemistry
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