Immunohistochemical Staining Protocols for Cancer Biomarkers

Six micrometer sections were obtained from the final CMA and TMA block for IHC stainings. p53 and EGFR immunohistochemical stainings was obtained from local clinical pathology department laboratory and carried out in Ventana staining automate. CIP2A immunohistochemical staining was carried out after protocol optimization in Ventana BenchMark XT staining automate (Ventana, Tucson, AZ) with OptiView DAB kit and with 64-min CC1 preparation and 32-min antibody incubation. Mouse monoclonal anti-CIP2A antibody (dilution 1:25, 2G10-3B5, sc-80,659, SantaCruz) was used. For PME-1, SET, LIMA1, NDFIP1, and Oct4 stainings, immunohistochemical stainings were done as previously described [13 (link), 24 (link)]. The antibodies used were rabbit polyclonal anti-SET (H-120) antibody sc-25,564 (diluted 1:1000, Santa Cruz Biotechnology), mouse monoclonal anti-PME-1 (B-12) antibody sc-25,278 (diluted 1:1000, Santa Cruz Biotechnology), rabbit polyclonal anti-NDFIP1 antibody HPA009682 (diluted 1:500, Sigma-Aldrich), and mouse monoclonal anti-Oct4 antibody sc5279 (diluted 1:200, Santa Cruz Biotechnology).

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Routila J., Suvila K., Grénman R., Leivo I., Westermarck J, & Ventelä S. (2021). Cancer cell line microarray as a novel screening method for identification of radioresistance biomarkers in head and neck squamous cell carcinoma. BMC Cancer, 21, 868.

Publication 2021

Ventana BenchMark XT OptiView DAB kit Mouse monoclonal anti-CIP2A antibody HPA009682 mouse monoclonal anti-Oct4 antibody

Anti antibody Antibodies Antibody Clinical laboratory Dilution Egfr Monoclonal antibody Mouse Oct4 Rabbit Stainings

Corresponding Organization :

Other organizations : Turku Centre for Computer Science, University of Turku, Åbo Akademi University, Turku University Hospital

Top 5 similar protocols

Variable analysis

independent variables

Immunohistochemical staining protocol optimization for CIP2A
Antibody dilutions for various proteins (p53, EGFR, CIP2A, SET, PME-1, NDFIP1, Oct4)

dependent variables

Immunohistochemical staining results for p53, EGFR, CIP2A, SET, PME-1, LIMA1, NDFIP1, Oct4

control variables

Tissue sections (six micrometer sections from final CMA and TMA blocks)
Ventana staining automate for immunohistochemical stainings
Previously described protocols for PME-1, SET, LIMA1, NDFIP1, and Oct4 stainings [13, 24]

positive controls

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negative controls

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