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Table 1.

Primary antibodies and detection methods

Antibody (clone)DilutionIncubation temperature and durationAntigen 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)DilutionIncubation temperature and durationAntigen 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.

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