Immunohistochemistry and Immunofluorescence Protocols

Immunohistochemistry (IHC) was performed as published previously [47 (link)] [7 (link)]. Tumor tissues were dissected, fixed in 4% formalin overnight and processed in paraffin wax. Signal detection was performed semiautomatically in the Autostainer 480 S (Medac, Hamburg, Germany). Nuclei were stained by hematoxylin. Immunofluorescence staining (IF) was performed as published [15 (link)] [47 (link)]. Nuclei were counterstained by Hoechst 33342. See S1 Table for antibody details and dilution ratios.

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Nettersheim D., Jostes S., Sharma R., Schneider S., Hofmann A., Ferreira H.J., Hoffmann P., Kristiansen G., Esteller M.B, & Schorle H. (2015). BMP Inhibition in Seminomas Initiates Acquisition of Pluripotency via NODAL Signaling Resulting in Reprogramming to an Embryonal Carcinoma. PLoS Genetics, 11(7), e1005415.

Publication 2015

Autostainer 480 S

Antibody Dilution Formalin Hoechst 33342 Immunofluorescence Immunohistochemistry Nuclei Paraffin wax Signal detection Tissues Tumor

Corresponding Organization : Universitat de Barcelona

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Protocol cited in 2 other protocols

Variable analysis

independent variables

Immunohistochemistry (IHC) protocol
Immunofluorescence staining (IF) protocol

dependent variables

Signal detection
Nuclei staining by hematoxylin
Nuclei counterstaining by Hoechst 33342

control variables

Tumor tissues dissected and fixed in 4% formalin overnight
Tumor tissues processed in paraffin wax
Signal detection performed semiautomatically in the Autostainer 480 S (Medac, Hamburg, Germany)

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