Table III.

Procedures for mutated p53, wild-type p53 and E6 staining.

Protocol	Mutated p53	p53 wild-type	E6
Blocking method	Reagent 1a; 5 min, RT	Reagent 1a; 5 min, RT	Reagent 1a; 5 min, RT
Primary antibody, dilution, incubation duration, incubation temperature, cat. no.	Anti-p53, 1:100 in PBS, 16 h, 4°C, ab32049b	Anti-p53, 1:200 in PBS, 16 h, 4°C, ab26b	Anti-E6, 1:150 in PBS, 1 h, RT, ab70b
Post blocking method	Reagent 2a; 20 min, RT	Reagent 2a; 20 min, RT	Reagent 2a; 20 min, RT
Secondary antibody, dilution, incubation duration, incubation temperature, cat. no.	HRP-Polymer Reagent 3a, 30 min, RT POLHRP-100	HRP-Polymer Reagent 3a, 30 min, RT POLHRP-100	HRP-Polymer Reagent 3a; 30 min, RT POLHRP-100
Chromogen	1 mg/ml DABc, 1 min	1 mg/ml DABc, 1 min	1 mg/ml DABc, 1 min

^aFrom the ZytoChem-Plus HRP Polymer-kit; Zytomed Systems GmbH, Berlin, Germany.

^bAbcam, Cambridge, UK.

^cDako, Glostrup, Denmark. RT, room temperature; HRP, horseradish peroxidase; DAB, 3,3′-diaminobenzidine.