Rabbit anti scp1

Meiotic cells were prepared as previously described by Massip et al. (2010) [25 (link)]. The synaptonemal complex proteins 3 (SCP3) and 1 (SCP1), MutL homolog 1 protein (MLH1) and centromeres were detected using the following primary antibodies: rabbit anti-SCP3 (1:1000; ABCAM, Cambridge, UK), rabbit anti-SCP1 (2:1000; ABCAM, Cambridge, UK), mouse anti-MLH1 (2:100; Becton Dickinson, Franklin Lakes, NJ), Human anti-kinetochore (1:100; Antibodies Incorporated, Davis), respectively, and prepared in a solution of PBT (PBS +0.16% BSA +0.1% Tween). The secondary antibodies consisted of Alexa 594 conjugated donkey anti-rabbit (1:100, Molecular Probes), Alexa 488 conjugated goat anti-mouse (1:100, Molecular Probes, Eugene, OR, USA), and AMCA conjugated donkey anti-human (1:100, Jackson Immunoresearch, West Grove, PA, USA). The γH2AX protein was detected by carrying out a complementary experiment without MLH1 antibody but with mouse anti-γH2AX (ABCAM, Cambridge, UK) and Alexa 488 conjugated goat anti-mouse (1:100, Molecular Probes, Eugene, OR, USA) antibodies. γH2AX is considered as a marker of unsynapsed chromatin (transcriptionally silenced chromosome regions—Turner et al. (2005) [26 (link)])

Meiotic cells were prepared as previously described by Massip et al. (2010) [31] . The synaptonemal complex proteins 3 (SCP3) and 1 (SCP1), MutL homolog 1 protein (MLH1) and centromeres were detected using the following primary antibodies: rabbit anti-SCP3 (1∶1000; ABCAM, Cambridge, UK), rabbit anti-SCP1 (2∶1000; ABCAM, Cambridge, UK), mouse anti-MLH1 (2∶100; Becton Dickinson, Francklin Lakes, NJ), Human anti-kinetochore (1∶100; Antibodysuits Incorporated, Davis), respectively, and prepared in a solution of PBT (PBS +0.16% BSA +0.1% Tween). The secondary antibodies consisted of Alexa 594 conjugated donkey anti-rabbit (1∶100, Molecular Probes), Alexa 488 conjugated goat anti-mouse (1∶100, Molecular Probes), and AMCA conjugated donkey anti-human (1∶100, Jackson, Grove, PA, USA). Spermatocytes were captured and analyzed using the Cytovision FISH imaging system (Leica Microsystems, Nanterre, France).