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4 protocols using anti dmc1

1

Immunofluorescence Staining of Germ Cell Markers

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The prepared sections were blocked with 3% (w/v) bovine serum albumin (BSA; ZSbio) in PBST (0.1% [v/v] Triton X-100 in PBS) for 1 h at room temperature and then incubated with the following primary antibodies overnight at 4°C: anti-Sycp3 (1:200, Abcam, Cambridge, MA, USA), anti-synaptonemal complex protein 1 (Sycp1; 1:200, Abcam), anti-Vasa (1:500, Abcam), anti-c-kit (1:500, Abcam), anti-Shp2 (1:200, Santa Cruz Biotechnology), anti-Plzf (1:500, Santa Cruz Biotechnology), anti-cleaved caspase 3 (1:200, Cell Signaling Technology, Boston, MA, USA), anti-Dmc1 (1:200, Abcam), anti-Smc3 (1:500, Abcam), and anti-DNA repair recombinase rad51 (Rad51; 1:200, Invitrogen). After being washed three times with PBST, the samples were incubated with the following secondary antibodies at a 1:200 dilution for 1 h at 37°C: Alexa Fluor 594/488-labeled anti-rabbit or anti-mouse IgG (YEASEN, Shanghai, China). The slides were subsequently washed three times in PBST and mounted with Vectashield containing 4’-6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA).
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2

Diverse Antibody Detection Protocols

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Anti-Flag (M2 Sigma or Wako), anti-HA (12CA5 for western blot, 16B12 for immunostaining, and F-7 for ChIP), anti-α-tubulin (Serotec), rabbit anti-Rad51 (Shinohara et al., 1992 (link)), anti-Dmc1 (Hayase et al., 2004 (link)), anti-Rad9 (Usui and Petrini, 2007 (link)), anti-Zip1 (Shinohara et al., 2008 (link)), and anti-H3K79-3me (Abcam) were used for western blot and immunostaining.
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3

Immunostaining of Meiotic Proteins

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For primary antibodies, we used goat antibody anti-SYCP3 (R&D Systems), mouse monoclonal antibody anti-DMC1 (Abcam ab11054), and a previously generated rabbit polyclonal anti-RAD51 [40 (link)]. For secondary antibodies, we used a donkey anti-rabbit IgG Alexa 488/647, donkey anti-mouse IgG Alexa 488/647, and donkey anti-goat Alexa 555 (Molecular Probes).
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4

Immunofluorescence Staining of Meiotic Proteins

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For primary antibodies, we used goat antibody anti-SYCP3 (R&D Systems), mouse monoclonal antibody anti-DMC1 (Abcam ab1837), and a previously generated rabbit polyclonal anti-RAD51 (38) . For secondary antibodies, we used a donkey anti-rabbit IgG . CC-BY 4.0 International license made available under a (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is
The copyright holder for this preprint this version posted January 6, 2020. ; https://doi.org/10.1101/2020.01.06.895680 doi: bioRxiv preprint 22 Alexa 488/647, donkey anti-mouse IgG Alexa 488/647, and donkey anti-goat Alexa 555 (Molecular Probes).
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