Immunohistochemistry and Immunofluorescence Protocols

IHC was performed as published previously (Nettersheim et
al, 2015). Tumour tissues were dissected, fixed in 4%
formalin overnight and processed in paraffin wax. Signal detection was performed
semi-automatically in the Autostainer 480S (Medac, Hamburg, Germany). Nuclei were
stained by hematoxylin. IF was performed as described previously (Nettersheim et al, 2011c (link), 2015 (link)). Nuclei were stained by Hoechst 33342. See Table 1 for antibody details and dilution ratios.

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Nettersheim D., Arndt I., Sharma R., Riesenberg S., Jostes S., Schneider S., Hölzel M., Kristiansen G, & Schorle H. (2016). The cancer/testis-antigen PRAME supports the pluripotency network and represses somatic and germ cell differentiation programs in seminomas. British Journal of Cancer, 115(4), 454-464.

Publication 2016

Autostainer 480S

Antibody Dilution Formalin Hematoxylin Hoechst 33342 Nuclei Paraffin wax Signal detection Tissues Tumour

Corresponding Organization :

Other organizations : University of Bonn

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Variable analysis

independent variables

IHC protocol
IF protocol

dependent variables

Signal detection
Nuclei staining

control variables

Tumor tissue dissection
Tissue fixation in 4% formalin
Paraffin wax processing
Semi-automatic signal detection in Autostainer 480S
Hoechst 33342 nuclei staining

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