Primary antibodies and detection methods
| Antibody (clone) . | Dilution . | Incubation temperature and duration . | Antigen retrieval buffer* . | Immunohistochemical detection method . |
|---|---|---|---|---|
| ATM (ATX08) | 1:25 | 4°C/20 h | CB | ImmunoMax† |
| BRCA1 (SG11) | 1:60 | 37°C/1 h | CB | ChemMate‡ |
| BRCA2§ | 1:500 | 4°C/20 h | CB | ImmunoMax† |
| Ku70§ | 1:1,000 | 37°C/0.5 h | CB | NBA-kit∥ |
| Ku80§ | 1:3,000 | 37°C/0.5 h | CB | NBA-kit∥ |
| MSH2 (FE11) | 1:50 | 37°C/1 h | AUS | EnVision‡ |
| MLH1 (G168-728) | 1:70 | 4°C/20 h | AUS | EnVision‡ |
| Ki67 (MIB 1) | 1:500 | 37°C/1 h | TEC | ChemMate‡ |
| Antibody (clone) . | Dilution . | Incubation temperature and duration . | Antigen retrieval buffer* . | Immunohistochemical detection method . |
|---|---|---|---|---|
| ATM (ATX08) | 1:25 | 4°C/20 h | CB | ImmunoMax† |
| BRCA1 (SG11) | 1:60 | 37°C/1 h | CB | ChemMate‡ |
| BRCA2§ | 1:500 | 4°C/20 h | CB | ImmunoMax† |
| Ku70§ | 1:1,000 | 37°C/0.5 h | CB | NBA-kit∥ |
| Ku80§ | 1:3,000 | 37°C/0.5 h | CB | NBA-kit∥ |
| MSH2 (FE11) | 1:50 | 37°C/1 h | AUS | EnVision‡ |
| MLH1 (G168-728) | 1:70 | 4°C/20 h | AUS | EnVision‡ |
| Ki67 (MIB 1) | 1:500 | 37°C/1 h | TEC | ChemMate‡ |
NOTE: Antibodies were supplied from Lab Vision Corp. Fremont, CA (ATM and BRCA2); Zymed Laboratories, San Francisco, CA (BRCA1); Oncogene Research Products, San Diego, CA (p53 and MSH2); BD Biosciences, San Jose, CA (MLH1); Dianova GmbH, Hamburg, Germany (Ki67); and DPC Biermann, Bad Nauheim, Germany (Ku70 and Ku80).
CB: 10 mmol/L citrate buffer, pH 6. AUS: ready-made antigen unmasking solution from Vector. TEC: 0.25 g Tris base, 0.5 g EDTA, 0.32 g citrate, pH 7.8.
For details of the ImmunoMax method, see ref. 34.
DakoCytomation Ltd., Ely, Cambridgeshire, United Kingdom.
Rabbit polyclonal antibody.
Zymed Laboratories.