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Immunohistochemistry: Paraffin Sections Using the Vectastain ABC Kit from Vector Labs
Published on: October 1, 2007
Immunohistochemistry: paraffin sections using the Vectastain ABC kit from vector labs
1Department of Physiology and Biophysics, University of California, Irvine, CA, USA. vchi@uci.edu
Journal of Visualized Experiments : Jove
|November 8, 2008
Summary
This study details indirect immunohistochemistry (IHC), a method using primary and secondary antibodies to visualize protein expression in tissue sections. The protocol covers paraffin-embedded slide preparation and staining for CD8 and tyrosine hydroxylase markers.
Area of Science:
- Biotechnology
- Immunology
- Histology
Background:
- Immunohistochemistry (IHC) visualizes protein expression in tissues using antibodies.
- Indirect IHC enhances signal and reduces background staining.
- Paraffin-embedded sections require specific preparation steps.
Purpose of the Study:
- To describe the indirect immunohistochemistry staining protocol.
- To detail the preparation of paraffin-embedded tissue sections for IHC.
- To demonstrate staining for CD8 and tyrosine hydroxylase.
Main Methods:
- Indirect IHC staining using primary and biotin-conjugated secondary antibodies.
- Preparation of paraffin-embedded sections: dewaxing, hydration, antigen retrieval, blocking.
- Staining for CD8 and tyrosine hydroxylase, followed by HRP-DAB development and counterstaining.
Main Results:
- Successful visualization of protein expression in tissue sections.
- Specific staining for CD8 and tyrosine hydroxylase markers achieved.
- Protocol yields slides ready for imaging.
Conclusions:
- Indirect IHC is an effective method for protein localization in tissues.
- The described protocol provides a reliable approach for preparing and staining paraffin-embedded sections.
- This technique is valuable for analyzing specific protein markers in biological samples.
