Four 3‐μm‐thick sections of FFPE samples were made for IHC analysis and stored at 37°C overnight. We used two auto‐staining systems. Protein expression was evaluated with mutL homolog 1 (MLH1) (M1; Ventana Medical Systems; Roche Group), mutS homolog 2 (MSH2) (G219‐1129; Ventana Medical Systems), mutS homolog 6 (MSH6) (44; Ventana Medical Systems), and postmeiotic segregation 1 homolog 2 (PMS2) (EPR3947; Ventana Medical Systems) antibodies using the Ventana BenchMark XT system (Roche). Protein expression was evaluated with MLH1 (ES05; Agilent), MSH2 (FE11; Agilent), MSH6 (EP49; Agilent), and PMS2 (EP51; Agilent) antibodies using the Autostainer Link48 (Agilent).27 , 28 All sections were evaluated by a medical technologist (K.A.) and a pathologist (T.O). Tumors with completely absent nuclear staining of at least one of MLH1, MSH2, MSH6, or PMS2 were classified as dMMR.
Msh2 g219 1129
Manufactured by Roche
The MSH2 (G219-1129) is a laboratory equipment product. It is a core component used in various scientific applications. The detailed specifications and intended use of this product are not available at this time.
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3 protocols using msh2 g219 1129
1
Immunohistochemical Analysis of MMR Proteins
2
Immunohistochemistry for Mismatch Repair Deficiency
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Immunohistochemistry was performed on formalin-fixed and paraffin-embedded tumor sections cut at 4-micron thickness and stained on a Ventana Bench Mark Autostainer (Ventana Medical System, Tucson, Arizona). The following rabbit monoclonal primary antibodies were used: MLH-1 (M1Ventana), PMS2 (EPR 3947, Ventana), MSH2 (G219-1129, Ventana), and MSH6 (44, Ventana). Antibodies were pre-diluted by the manufacturer and staining was performed following the manufacturer’s protocols in collaboration with platform vendors. Technical methodologies and quality assurance were performed at the Immunopathology Laboratory of Long Island Jewish Medical Center (Northwell Health System, New Hyde Park, NY). Complete loss of any of the foregoing markers lead to the designation of MMR deficient.
3
Immunohistochemical Evaluation of MMR Status
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Immunohistochemistry was performed in order to determine MMR status on formalin-fixed and paraffin-embedded tumor sections cut at 4-μM thickness and stained on a Ventana Bench Mark Autostainer (Ventana Medical System, Tucson, AZ). The following rabbit monoclonal pre-diluted primary antibodies were used: MLH-1(M1, Ventana), PMS2 (EPR 3947, Ventana), MSH2(G219-1129, Ventana), and MSH6 (44, Ventana). Technical methodologies and quality assurance were undertaken by certified histotechnologists at the immunohistochemical laboratory of Long Island Jewish Medical Center (Northwell Health System, New Hyde Park, NY). Cases showing loss of immunohistochemical staining (<1%) for any of the following stains: MLH1, PMS2, MSH2, and MSH6 were considered MMR-deficient.
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