Surface-spread chromosomes of mouse spermatocytes were prepared from 18 days postpartum male mice using the drying-down technique as previously described33 (link). Immuno-staining was performed as previously described33 (link) using the following primary and secondary antibodies: goat anti-SYCP3 (Santa Cruz Biotechnology, sc-20845; 1:200), rabbit anti-MSH4 (a gift from Dr. Paula Cohen; 1:200), mouse monoclonal anti-MLH1 (4C9C7) (Cell Signaling Technology, 3515S; 1:30), AMCA anti-goat (Jackson ImmunoResearch, 705–155-147; 1:50), Cy3 anti-rabbit (Jackson ImmunoResearch, 711–165-152; 1:100), and AlexaFluor647 anti-mouse (Jackson ImmunoResearch, 715–605-151; 1:100).
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Immunostaining of Mouse Spermatocyte Chromosomes
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Corresponding Organization :
Other organizations : Howard Hughes Medical Institute, University of California, Davis
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Variable analysis
independent variables
- Age of male mice (18 days postpartum)
- SYCP3 (synaptonemal complex protein 3) expression
- MSH4 (MutS Homolog 4) expression
- MLH1 (MutL Homolog 1) expression
- Drying-down technique for preparing surface-spread chromosomes of mouse spermatocytes
- Immuno-staining procedure for SYCP3, MSH4, and MLH1
- Primary and secondary antibodies used for immuno-staining
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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