| Antibody . | Dilution . | Source . | Conditions . |
|---|---|---|---|
| Immunofluorescence | |||
| Mouse anti-pH3-T3 | 1:10,000 | Gift, Dr. Hiroshi Kimura (Kelly et al., 2010) | 1% TX 3 min, 4% PFA 20 min |
| Mouse anti-Sgo1 | 1:500 | Abcam 58023 | 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-ABK | 1:500 | Abcam 2254 | 0.5% TX 30 s, 4% PFA 20 min; 1% TX 3 min, 4% PFA 20 min |
| Mouse anti-Bub1 | 1:200 | Abcam 54893 | 0.5% TX 30 s, 4% PFA 20 min; 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-Bub1 | 1:200 | Bethyl Laboratories A300-373A | 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-pH2A-T120 | 1:1,000 | Active Motif 39391 | 0.5% TX 30 s, 4% PFA 20 min; 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-pABK-T232 | 1:500 | (DeLuca et al., 2017) | 0.5% TX 30 s, 4% PFA 20 min; 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-pDsn1-S109 | 1:500 | Gift, Dr. Iain Cheeseman (Welburn et al., 2010) | 0.5% TX 30 s, 4% PFA 20 min |
| Rabbit anti-pHec1-S44 | 1:1,000 | (DeLuca et al. 2011) | 0.5% TX 30 s, 4% PFA 20 min; 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-pINCENP-S893,S894 | 1:400 | This study | 0.5% TX 30 s, 4% PFA 20 min |
| Rabbit anti-pKNL1-S24 | 1:400 | Gift, Dr. Iain Cheeseman (Welburn et al., 2010) | 0.5% TX 30 s, 4% PFA 20 min |
| Rabbit anti-Survivin | 1:500 | Santa Cruz Biotechnology 10811 | 1% TX 3 min, 4% PFA 20 min |
| Two-color distance measurements for centromere/kinetochore mapping | |||
| Mouse anti-Hec1 (9G3) | 1:2,000 | GeneTex GTX70268 | 0.5% TX 30 s, 4% PFA 20 min |
| Guinea pig anti-CENP-C | 1:500 | MBL PD030 | 0.5% TX 30 s, 4% PFA 20 min |
| Rabbit anti-pH2A-T120 | 1:1,000 | Active Motif 39391 | 0.5% TX 30 s, 4% PFA 20 min |
| Rabbit anti-pABK-T232 | 1:500 | (DeLuca et al., 2017) | 0.5% TX 30 s, 4% PFA 20 min |
| Western blotting | |||
| Rabbit anti-pABK-T232 | 1:500 | (DeLuca et al., 2017) | |
| Rabbit anti-pINCENP-S893,S894 | 1:500 | This study | |
| Antibody . | Dilution . | Source . | Conditions . |
|---|---|---|---|
| Immunofluorescence | |||
| Mouse anti-pH3-T3 | 1:10,000 | Gift, Dr. Hiroshi Kimura (Kelly et al., 2010) | 1% TX 3 min, 4% PFA 20 min |
| Mouse anti-Sgo1 | 1:500 | Abcam 58023 | 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-ABK | 1:500 | Abcam 2254 | 0.5% TX 30 s, 4% PFA 20 min; 1% TX 3 min, 4% PFA 20 min |
| Mouse anti-Bub1 | 1:200 | Abcam 54893 | 0.5% TX 30 s, 4% PFA 20 min; 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-Bub1 | 1:200 | Bethyl Laboratories A300-373A | 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-pH2A-T120 | 1:1,000 | Active Motif 39391 | 0.5% TX 30 s, 4% PFA 20 min; 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-pABK-T232 | 1:500 | (DeLuca et al., 2017) | 0.5% TX 30 s, 4% PFA 20 min; 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-pDsn1-S109 | 1:500 | Gift, Dr. Iain Cheeseman (Welburn et al., 2010) | 0.5% TX 30 s, 4% PFA 20 min |
| Rabbit anti-pHec1-S44 | 1:1,000 | (DeLuca et al. 2011) | 0.5% TX 30 s, 4% PFA 20 min; 1% TX 3 min, 4% PFA 20 min |
| Rabbit anti-pINCENP-S893,S894 | 1:400 | This study | 0.5% TX 30 s, 4% PFA 20 min |
| Rabbit anti-pKNL1-S24 | 1:400 | Gift, Dr. Iain Cheeseman (Welburn et al., 2010) | 0.5% TX 30 s, 4% PFA 20 min |
| Rabbit anti-Survivin | 1:500 | Santa Cruz Biotechnology 10811 | 1% TX 3 min, 4% PFA 20 min |
| Two-color distance measurements for centromere/kinetochore mapping | |||
| Mouse anti-Hec1 (9G3) | 1:2,000 | GeneTex GTX70268 | 0.5% TX 30 s, 4% PFA 20 min |
| Guinea pig anti-CENP-C | 1:500 | MBL PD030 | 0.5% TX 30 s, 4% PFA 20 min |
| Rabbit anti-pH2A-T120 | 1:1,000 | Active Motif 39391 | 0.5% TX 30 s, 4% PFA 20 min |
| Rabbit anti-pABK-T232 | 1:500 | (DeLuca et al., 2017) | 0.5% TX 30 s, 4% PFA 20 min |
| Western blotting | |||
| Rabbit anti-pABK-T232 | 1:500 | (DeLuca et al., 2017) | |
| Rabbit anti-pINCENP-S893,S894 | 1:500 | This study | |
For each antibody used for standard immunofluorescence experiments, two-color distance measurements, and Western blots, the table indicates the concentration used, antibody source, and fixation conditions. TX, Triton X-100; PFA, paraformaldehyde.