Table 1. Summary of double-IFa and triple-IFb staining methods.
| cyclin B1 | ||||
|---|---|---|---|---|
| Antibody staining | Separase double- and triple-IF | Securin double- and triple-IF | Double-IF | Triple-IF |
| Type | Mouse monoclonal | Rabbit monoclonal | Rabbit monoclonal | mouse monoclonal |
| Clone | 6H6 | EPR3240 | Y106 | V152 |
| Source | Novus Biologicals H00009700-M01 | Abcam ab79546 | Abcam ab32053 | MA5-13128 |
| Dilution | 1:100 or 1:1600c | 1:500 1h RT | 1:100 1h RT | 1:600 1h RT |
| Antibody | Ki-67 | PHH3 | Cleaved caspase-3 | |
| Staining | double-IF | double-IF | double-IF | |
| Type | Rabbit polyclonal | Rabbit polyclonal | Rabbit monoclonal | |
| Clone | Polyclonal | Polyclonal | D3E9 | |
| Source | Chemicon International AB9260 | Cell Marque 369A-14 | Cell Signaling Technology #9579 | |
| Dilution | 1:1000 | 1:100 | 1:100 | |
a
Tyramide Signal Amplification HRP Kit, Molecular Probes by LifeTechnologies, Eugene, OR, USA.
b
Goat Anti-Mouse IgG2a Human ads-HRP, 1080-05, SouthernBiotech, Birmingham, AL, USA. Tyramide Signal Amplification HRP Kit, Molecular Probes by LifeTechnologies, Eugene, OR, USA. Goat Anti-Mouse IgG1 Human ads-HRP, 1070-05, SouthernBiotech, Birmingham, AL, USA.
c
Dilutions and incubations times and temperatures were optimised for each lot of antibody.