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Updated: Mar 29, 2026
Indirect Immunofluorescence on Frozen Sections of Mouse Mammary Gland
Published on: December 1, 2015
Indirect Immunofluorescence on Frozen Sections of Mouse Mammary Gland
Edith Honvo-Houéto1, Sandrine Truchet2
1Jouy-en-Josas Centre, INRA.
Insights
This study details a straightforward indirect immunofluorescence protocol for detecting and localizing proteins within tissues. The method covers tissue preparation, antibody staining, and image analysis, offering a comprehensive guide for cellular protein localization.
Area of Science:
- Biotechnology
- Cell Biology
- Immunology
Background:
- Indirect immunofluorescence is a key technique for protein detection and localization in biological tissues.
- Standardized protocols are essential for reproducible results in cellular and molecular research.
Purpose of the Study:
- To present a complete and simplified protocol for indirect immunofluorescence.
- To demonstrate the application of the protocol using the mouse lactating mammary gland as an example.
- To provide a comprehensive overview from tissue collection to protein localization.
Main Methods:
- Detailed tissue preparation including dissection, fixation, and frozen sectioning.
- Standard indirect immunofluorescence staining with an optional antigen retrieval step.
- Image acquisition and post-treatment for labeled tissue sections.
Main Results:
- A reproducible method for indirect immunofluorescence was established.
- Successful cellular localization of proteins in mouse mammary gland tissue was achieved.
- The protocol provides a clear workflow from tissue handling to protein visualization.
Conclusions:
- The presented protocol offers a user-friendly approach for protein detection and cellular localization via indirect immunofluorescence.
- The method is adaptable to various tissue types and primary antibodies with appropriate adjustments.
- This technique is valuable for researchers studying protein expression and distribution in diverse biological systems.
Abstract:
Indirect immunofluorescence is used to detect and locate proteins of interest in a tissue. The protocol presented here describes a complete and simple method for the immune detection of proteins, the mouse lactating mammary gland being taken as an example. A protocol for the preparation of the tissue samples, especially concerning the dissection of mouse mammary gland, tissue fixation and frozen tissue sectioning, are detailed. A standard protocol to perform indirect immunofluorescence, including an optional antigen retrieval step, is also presented. The observation of the labeled tissue sections as well as image acquisition and post-treatments are also stated. This procedure gives a full overview, from the collection of animal tissue to the cellular localization of a protein. Although this general method can be applied to other tissue samples, it should be adapted to each tissue/primary antibody couple studied.
