Immunohistochemistry was performed on formalin-fixed paraffin-embedded tissue using 4 μm sections cut from paraffin blocks. Staining was performed using a fully automated system (Benchmark ULTRA, Ventana Medical Systems, Tuscon, AZ, USA) and the following antibodies were used: NTRK1 (Ab76291, 1:1,500, ABCAM), SOX10 (383A-75,1:50, CELL MARQUE), S100 (Z0311, 1:8,000, DAKO), H3 (9733, 1:100, CELL SIGNALING), CD34 (790-2927, VENTANA), SMA (VPS281, 1:50, VECTOR), Desmin (760-2513, VENTANA), Melan A (790-2990, VENTANA), STAT6 (SC-621, 1:2,500, SANTA CRUZ) and HMB45 (M0634, 1:100, DAKO).
Melan a
Manufactured by Roche
Sourced in United States
Melan A is a lab equipment product designed for the detection and quantification of the Melan A protein, which is commonly used as a biomarker in histopathological examinations. The core function of this product is to facilitate the identification and analysis of Melan A levels in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Sox10, by Cell Marque (2 mentions)
Hmb 45, by Roche (2 mentions)
Ab76291, by Abcam (1 mentions)
Desmin, by Roche (1 mentions)
Hmb45, by Agilent Technologies (1 mentions)
M0634, by Agilent Technologies (1 mentions)
Benchmark ultra, by Roche (1 mentions)
Ventana benchmark ultra, by Roche (1 mentions)
At2 whole slide scanner, by Leica (1 mentions)
4 protocols using melan a
1
Comprehensive Immunohistochemical Profiling
2
Retrospective Study of Primary Cutaneous Melanoma
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This retrospective study was conducted in the Department of Pathology of the County Clinical Hospital Mureș between January 2021 and June 2022 and included all cases of PCM diagnosed in this department beginning in 2015. The criteria for inclusion in the study were invasive melanoma with complete IHC profile (S-100, SOX 10, HMB-45, Melan-A). The following exclusion criteria were applied: metastatic or desmoplastic melanoma, inadequate depth of the biopsy specimen, local re-excisions, PCMs associated with a nevus, and uninterpretable IHC reactions. The study was conducted in accordance with the Declaration of Helsinki and was approved by the Ethics Committee of the hospital (protocol number 16381/06.01.2021). For each case, five slides with 3 µm thickness were selected, stained with HE, S-100 (polyclonal antibody, Ventana, AZ, USA), SOX10 (SP267, primary monoclonal antibody, Cell Marque, CA, USA), HMB-45 (monoclonal antibody, Ventana, AZ, USA), and Melan-A (monoclonal antibody, Ventana, AZ, USA). Furthermore, all sections had previously been obtained from the same paraffin block. The process of case selection is illustrated in Figure 1 .
3
Lymph Node Histopathology Examination
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For histopathological examination, standard staining consisting of haematoxylin and eosin staining was performed. In addition, immunohistochemical workup of lymph node tissue was performed according to the underlying tumour type on an automated platform (Ventana Benchmark Ultra, Ventana Medical Systems, AZ, USA) adherent to manufacturer's protocols as follows: Melan-A (Clone: A103, ready to use kit; Ventana Medical Systems) and MITF (Clone: D5, 1:100; Agilent Dako, CA, USA) for melanoma and CK20 (Clone: Ks20.8, 1:1000; Agilent Dako) and Synaptophysin (Clone: MRQ-40, ready to use kit; Ventana Medical Systems) for Merkel cell carcinoma. Histopathologic evaluation of the lymph nodes was performed by an experienced pathologist from the Institute of Pathology or by an experienced dermatopathologist from the Department of Dermatology, Essen University Hospital. Histopathological lymph node specimens were digitised with the Aperio AT2 whole-slide scanner (Leica, Wetzlar, Germany) at 20× resolution.
4
Histological Analysis of Sentinel Lymph Nodes
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All harvested SLNs were fixed in formalin, serially sectioned at 2 mm intervals, and submitted in cassettes for paraffin embedding. Unstained slides were cut at 40 µm intervals for preparation of two flanking hematoxylin and eosin (H&E) slides and four slides for immunohistochemical stains (Melan A, HMB45, S100, and negative control; Ventana Medical Systems, Tucson, AZ, USA; Benchmark Ultra Stainer) for histologic examination. All harvested non-SLNs were sectioned once for H&E staining only.
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