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Updated: Jul 6, 2026
Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing
Published on: March 15, 2019
Comparative analysis of anti-mouse SRY antibodies
S T Bradford1, D Wilhelm, P Koopman
1Division of Molecular Genetics and Development, The University of Queensland, Brisbane, Qld, Australia.
Abstract:
The Y chromosome gene SRY is the initiator of male sexual differentiation in mammals, but the molecular and cellular mechanisms operating downstream of SRY remain undefined. A deeper understanding of these issues relies on the ability to visualize SRY protein endogenously under a number of experimental conditions. Here we compare the specificity and effectiveness of several available antibodies to mouse SRY. Two antibodies cross-reacted with other SOX proteins in immunofluorescence analyses of transfected cells, and one of these two was unable to detect SRY on Western blots. A third antibody was both avid and specific, and was able to detect endogenous SRY in developing Sertoli cells in mouse genital ridges. Our findings underline the need to distinguish between useful and spurious reagents for biochemical and immunolocalization studies involving mouse SRY protein.
Insights
Identifying reliable antibodies is crucial for studying the SRY gene, the key initiator of male sex determination. This research evaluates mouse SRY antibodies, finding one specific antibody effective for detecting endogenous SRY protein in Sertoli cells.
Area of Science:
- Genetics
- Developmental Biology
- Molecular Biology
Background:
- The Y chromosome gene SRY initiates male sexual differentiation in mammals.
- Molecular mechanisms downstream of SRY are not fully understood.
- Visualizing endogenous SRY protein is essential for studying its function.
Purpose of the Study:
- To compare the specificity and effectiveness of available antibodies for mouse SRY.
- To identify reliable reagents for studying SRY protein.
- To enable visualization of endogenous SRY in relevant cellular contexts.
Main Methods:
- Immunofluorescence analysis of transfected cells with different antibodies.
- Western blot analysis to assess antibody detection of SRY.
- Immunolocalization studies in developing mouse genital ridges.
Main Results:
- Two tested antibodies showed cross-reactivity with other SOX proteins.
- One antibody failed to detect SRY on Western blots.
- A third antibody demonstrated high specificity and avidity, detecting endogenous SRY in Sertoli cells.
Conclusions:
- Not all commercially available antibodies are suitable for mouse SRY research.
- A specific and avid antibody is identified for detecting endogenous SRY.
- Validated reagents are necessary for accurate biochemical and immunolocalization studies of SRY protein.
