Validated Immunostaining Antibodies for C. elegans

The following primary antibodies were used at the indicated dilutions: rabbit anti-GFP-488-conjugated (1:200) (Invitrogen, A21311), goat anti-GFP-FITC-conjugated (1:200) (Abcam, ab6662), rat anti-mCherry (1:1000) (Chromotek, 5F8), rabbit anti-COH-3/4 (1:400)^{21 (link)}, mouse anti-REC-8 (1:100) (Novus Biologicals, 29470002), guinea pig anti-SYP-1 (1:400)^{26 (link)}, chicken anti-SYP-1 (1:300)^{47 (link)}, guinea pig anti-HTP-3 (1:800)^{32 (link)}, rabbit anti-HIM-3 (1:400)^{33 (link)}, rabbit anti-HTP-1/2 (1:400)^{47 (link)}, rabbit anti-PLK-2 (1:500)^{65 (link)}, guinea pig anti-SUN-1 pS12 (1:1000)^{19 (link)}, rabbit anti-RAD-51 (1:10000) (Novus Biologicals, 29480002), mouse anti-HA (1:200) (Cell Signalling, 23675), rabbit anti-DSB-2 (1:1000)^{9 (link)}, rabbit anti-HIM-8 (1:500) (Novus Biologicals, 41980002). Phospho-specific antibodies against HIM-8 pT64 (1:400) were produced by injecting rabbits with the synthetic peptide DTPRFSpTPIVPNVC (GenScript). Polyclonal anti-HIM-8 pT64 antibodies were affinity purified by binding to a column containing the phospho-peptide and the specificity of the antibodies was validated by absence of staining in germlines of chk-2 mutant worms.