Tissue Microarray Immunohistochemistry for p53

The construction and analysis of tissue microarray containing all these human samples has been reported elsewhere^{4 (link),47 (link)}. At least two representative duplicate cores for each case were scored. Inmunohistochemistry analysis was carried out using the monoclonal mouse anti-human p53 (DO07, Agilent-Dako). Signal was amplified using avidin-peroxidase (ABC Elite Kit; Vector Labs), and peroxidase was visualized using 3,30-diaminobenzidine as a substrate (DAB kit, Vector Labs). Negative control slides were obtained by replacing primary antibodies with PBS (data not shown). Scoring of the results and selection of the thresholds, internal controls for reactivity of each antibody, and tissue controls for the series were done according to previously published methods^{4 (link),47 (link)}.

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Dueñas M., Pérez-Figueroa A., Oliveira C., Suárez-Cabrera C., Sousa A., Oliveira P., Villacampa F., Paramio J.M, & Martínez-Fernández M. (2019). Gene Expression Analyses in Non Muscle Invasive Bladder Cancer Reveals a Role for Alternative Splicing and Tp53 Status. Scientific Reports, 9, 10362.

Publication 2019

ABC Elite Kit DAB kit

Antibodies Antibody Avidin Human Mouse Peroxidase Tissue Tissue microarray analysis Vector

Corresponding Organization :

Other organizations : Universidad Complutense de Madrid, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas, Universidade de Vigo, Universidade do Porto, Centro de Investigación Biomédica en Red de Cáncer

Top 5 similar protocols

Variable analysis

independent variables

Immunohistochemistry analysis using the monoclonal mouse anti-human p53 (DO07, Agilent-Dako) antibody

dependent variables

P53 protein expression

control variables

Tissue microarray containing all the human samples
At least two representative duplicate cores for each case were scored
Signal was amplified using avidin-peroxidase (ABC Elite Kit; Vector Labs)
Peroxidase was visualized using 3,30-diaminobenzidine as a substrate (DAB kit, Vector Labs)
Negative control slides were obtained by replacing primary antibodies with PBS

positive controls

Internal controls for reactivity of each antibody
Tissue controls for the series

negative controls

Negative control slides were obtained by replacing primary antibodies with PBS

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