Immunohistochemical Detection of HPyV7 LTag in Thymomas

PRb and phosphoralized Rb (phospho-Rb) expression was detected by using two monoclonal Retinoblastoma antibodies: pRb (a.a. 332–344), clone G3-245, Pharmingen, dilution 1:300 and phospho-Rb, clone D20B12, dilution 1:100. P16 expression was performed with a monoclonal antibody (clone JC8, dilution 1:400) (Santa Cruz). Secondary antibody detection and visualization were done with the EnVision FLEX™ Kit K8008 (DAKO) or K5005 (Dako) according to standard protocols. Expression levels were assessed and scored by three experienced investigators (AzH, MAH, DR). The results of the HPyV7 LTag expression using the 2 t10 antibody have been described previously [6 ]. In the epithelial cells of 17 thymomas (46 %) marked LTag expression was found. The expression of LTag was in good agreement with earlier performed HPyV7-DNA PCR and/or the HPyV7-FISH [7 (link)].

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Keijzers M., Rensspiess D., Pujari S., Abdul-Hamid M.A., Hochstenbag M., Dingemans A.M., Kurz A.K., Haugg A., Maessen J.G., De Baets M.H, & zur Hausen A. (2015). Expression of pRb and p16INK4 in human thymic epithelial tumors in relation to the presence of human polyomavirus 7. Diagnostic Pathology, 10, 201.

Publication 2015

clone G3-245

Antibody Clone Dilution Epithelial cells Fish Monoclonal antibody Retinoblastoma Thymomas

Corresponding Organization :

Other organizations : Maastricht University Medical Centre, Universitätsklinikum Aachen, Maastricht University

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Variable analysis

independent variables

PRb antibody (a.a. 332–344), clone G3-245, Pharmingen, dilution 1:300
Phospho-Rb antibody, clone D20B12, dilution 1:100
P16 antibody, clone JC8, dilution 1:400 (Santa Cruz)

dependent variables

PRb expression
Phospho-Rb expression
P16 expression
HPyV7 LTag expression

control variables

Secondary antibody detection and visualization with EnVision FLEX™ Kit K8008 (DAKO) or K5005 (Dako) according to standard protocols
Expression levels assessed and scored by three experienced investigators (AzH, MAH, DR)

positive controls

The results of the HPyV7 LTag expression using the 2 t10 antibody have been described previously [6 (no link found)]

negative controls

None mentioned

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