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Stripping of the Immunoblot for Reprobing

Authors: Larisa, Litovchick;

Stripping of the Immunoblot for Reprobing

Abstract

For most immunoblots developed with chemiluminescence or with fluorochrome-based detection systems, it is possible to remove the primary and secondary antibodies from the membrane without affecting the bound antigen. This allows you to reuse the membrane for detection of another protein antigen. The blots developed with chromogenic substrates can also be stripped of antibodies and reprobed, but the bands detected in the first round of immunoblotting will remain unaffected. Stripping and reprobing of the membrane are particularly useful when the amount of sample is limited or when it is important to accurately compare the signal between two different protein antigens in the same sample. Examples of such experiments include determining the levels of a protein antigen in a series of samples relative to the loading control and comparison of the phosphorylated form to the total levels of the protein in the sample.

Keywords

Staining and Labeling, Immunoblotting, Collodion, Proteins, Membranes, Artificial, Buffers, Antibodies, Fluorocarbon Polymers, Molecular Probes, Electrophoresis, Polyacrylamide Gel, Polyvinyls, Antigens, Fluorescent Dyes

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Powered by OpenAIRE graph
Found an issue? Give us feedback
selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
10
Top 10%
Average
Top 10%
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