Chapter title |
Immunofluorescence Microscopy for DIGE-Based Proteomics
|
---|---|
Chapter number | 23 |
Book title |
Difference Gel Electrophoresis
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7268-5_23 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7267-8, 978-1-4939-7268-5
|
Authors |
Rustam R. Mundegar, Margit Zweyer, Dieter Swandulla |
Abstract |
Alterations in the proteome of a tissue in different settings, as assessed by difference gel electrophoresis, can be verified for single proteins using immunohistochemistry. In fluorescence immunohistochemistry, an antibody to a particular antigen is applied to tissue sections, and fluorophores conjugated to a secondary antibody allow for the detection of target antigen with fluorescent microscopy. Visual comparison is sufficient for the detection of significant alterations in the abundance of a certain protein in different settings. Additionally, unlike large-scale proteome analyses and Western blot methods, expression of target protein can be analyzed at the cellular level by immunohistochemistry. In this chapter, a protocol for the application of fluorescence immunohistochemistry for the detection of dystrophin in skeletal muscle sections is outlined, including sample preparation, tissue sectioning, and immunostaining. |
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