Pan-TRK IHC was conducted using the VENTANA pan-TRK (EPR17341) Assay (Roche Diagnostics, Basel, Switzerland) per the manufacturer protocol with the following modifications: staining was performed on a Leica Bond III platform (Leica Instruments, Wetzlar, Germany). Antigen retrieval at pH 9.0 was performed for 20 min. Incubation time with the primary pan-TRK antibody (EPR17341) was at room temperature for 15 min. Detection was performed using Polymer Refine Detection System (Leica) and 3,3สน-diaminobenzidine chromagen. On slide controls for IHC was cerebellum (membranous and cytoplasmic staining in neurons).
Ventana pan trk epr17341 assay
Manufactured by Roche
Sourced in Switzerland, Italy
The VENTANA pan-TRK (EPR17341) Assay is a laboratory equipment product developed by Roche. It is an immunohistochemistry (IHC) assay designed to detect the presence of pan-TRK proteins in tissue samples. The assay utilizes the EPR17341 antibody clone to identify cells expressing TRK proteins, which are involved in various cellular processes.
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2 protocols using ventana pan trk epr17341 assay
1
Pan-TRK IHC Protocol on Leica Bond III
2
Immunohistochemical Detection of Trk Proteins
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For each of the included cases, a 4 um-thick slice was obtained from FFPE tissue blocks for IHC analysis; the most representative tissue block, the same as previously employed for NTRK fusion detection, was used. The VENTANA pan-TRK (EPR17341) assay (Roche Diagnostics Spa, Monza, MB, Italy) was used to assess the expression of Trk proteins. In detail, this in vitro-validated assay enables the detection of C-terminal region of TrkA, TrkB and TrkC, which should be maintained in case of NTRK1, NTRK2 and NTRK3 rearrangements. All procedures were conducted according to the methodology protocol. A positive control (appendix tissue) was included in each experimental session, as recommended by the manufacturer.
IHC staining was interpreted by three qualified pathologists. Positivity was deemed in cases with signal above background in at least 1% of tumor cells, as indicated by the manufacturer. Signal intensity, percentage of positive cells and subcellular staining localization (membranous, cytoplasmic and nuclear) were recorded. In discordant cases, a consensus was reached by collegial discussion. Signal intensity was expressed as a score, from 1 to 3, corresponding to weak, mild and strong signals [31 (link)].
IHC staining was interpreted by three qualified pathologists. Positivity was deemed in cases with signal above background in at least 1% of tumor cells, as indicated by the manufacturer. Signal intensity, percentage of positive cells and subcellular staining localization (membranous, cytoplasmic and nuclear) were recorded. In discordant cases, a consensus was reached by collegial discussion. Signal intensity was expressed as a score, from 1 to 3, corresponding to weak, mild and strong signals [31 (link)].
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