Clone g219 1129

Manufactured by Roche

Sourced in Switzerland

Clone G219-1129 is a laboratory equipment product. It serves as a core function for its intended use, but a detailed unbiased description is not available at this time.

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G219-1129 (15 citations) MSH2 (clone G219-1129, (13 citations)

8 protocols using clone g219 1129

Immunohistochemical Analysis of Formalin-Fixed Paraffin-Embedded Tissue

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Formalin-fixed paraffin-embedded tissue was collected from the archives of the pathology division of our hospital. Hematoxylin–eosin archival slides were revised by an expert pathologist and the diagnosis was confirmed in all cases. The most representative slide of each sample was selected for IHC in cases with available material. Specifically, three-micron-thick serial paraffin sections of each case were processed by IHC using an automated immunostainer (Ventana BenchMark Ultra AutoStainer, Ventana Medical Systems, Tucson, AZ, USA) with antibodies against p53 (clone DO-7, catalogue number 7800-2912,Ventana Medical Systems, Tucson, AZ, USA), ER (clone SP1, catalogue number 790-4324, Ventana Medical Systems, Tucson, AZ, USA), PgR (clone 1E2, catalogue number 790-2223, Ventana Medical Systems, Tucson, AZ, USA), and the MMR status, evaluating the protein expression of MLH1 (clone M1, catalogue number 760-5091, Ventana Medical Systems, Tucson, AZ, USA), PMS2 (clone EPR3947, catalogue number 760-5094, Ventana Medical Systems, Tucson, AZ, USA), MSH2 (clone G219-1129, catalogue number 760-5093, Ventana Medical Systems, Tucson, AZ, USA), MSH6 (clone 44, catalogue number 760-5092, Ventana Medical Systems, Tucson, AZ, USA). Appropriate positive controls were included for each IHC run.

Bounous V.E., Ferrero A., Campisi P., Fuso L., Pezua Sanjinez J.O., Manassero S., De Rosa G, & Biglia N. (2022). Immunohistochemical Markers and TILs Evaluation for Endometrial Carcinoma. Journal of Clinical Medicine, 11(19), 5678.

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Tumor Imaging and MMR Status Assessment

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Tumor imaging was conducted at baseline, every 8 weeks until the primary analysis for the study, and then every 12 weeks thereafter. Response was assessed by blinded independent central review (BICR) per RECIST version 1.1. Adverse events (AEs) were graded per the National Cancer Institute Common Terminology Criteria for Adverse Events, version 4.0, and were monitored throughout the study and for 30 days (120 days for serious AEs) after treatment discontinuation.
Tumor tissue was collected from all enrolled patients for determination of MMR status by central assessment by pathologist evaluation before randomization. Mismatch repair was assessed in archived tumor tissue from the most recent surgery/biopsy or from a fresh biopsy if no archival tumor tissue was available. Automated immunohistochemistry staining and chromogenic labeling of the MMR proteins MLH1, MSH2, MSH6, and PMS2 on the Ventana Benchmark Ultra was performed using mouse and rabbit antibodies (Roche Diagnostics). Specifically, the MLH1 (clone M1, mouse monoclonal, Ventana, Cat# 790–5091), PMS2 (clone A16‐4, mouse monoclonal, Ventana, Cat# 790–5094), MSH2 (clone G219‐1129, mouse monoclonal, Ventana, Cat# 790–5093), and MSH6 (clone SP93, rabbit monoclonal, Ventana, Cat# 790–5092) antibodies were used to perform immunohistochemistry staining.

Yonemori K., Yunokawa M., Ushijima K., Sakata J., Shikama A., Minobe S., Usami T., Enomoto T., Takehara K., Hasegawa K., Yamagami W., Yamamoto K., Han S., Dutta L., Orlowski R., Miura T., Makker V, & Fujiwara K. (2022). Lenvatinib plus pembrolizumab in Japanese patients with endometrial cancer: Results from Study 309/KEYNOTE‐775. Cancer Science, 113(10), 3489-3497.

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Immunohistochemical Evaluation of MMR Status

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MMR status was evaluated independently, on full stained slides using Ready-to-Use monoclonal antibodies against MLH1 (clone M1, Ventana/Roche, Basal, Schweiz), MSH6 (clone SP93, Ventana/Roche, Basal, Schweiz), MSH2 (clone G219-1129, Ventana/Roche, Basal, Schweiz) and PMS2 (clone A16-4, Ventana/Roche, Basal, Schweiz).

Choo J., Kua L.F., Soe M.Y., Asuncion B.R., Tan B.K., Teo C.B., Tay R.Y., So J., Shabbir A., Guowei K., Tan H.L., Chan G., Ma H., Ramachandran G.K., Lum J.H., Chee C.E., Sridharan S., Tan P., Sundar R, & Yong W.P. (2023). Clinical relevance of PD-1 positive CD8 T-cells in gastric cancer. Gastric Cancer, 26(3), 393-404.

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Mismatch Repair Protein Immunohistochemistry

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Formalin-fixed, paraffin-embedded tumors were stained for MLH1, MSH2, MSH6, and PMS2 proteins. Mismatch repair protein loss is defined as the absence of nuclear staining in neoplastic cells but positive nuclear staining in lymphocytes and normal adjacent colonic epithelium (16 (link)). Primary monoclonal antibodies against MLH1 (clone M1, Ventana, prediluted), MSH2 (clone G219-1129, Ventana, prediluted), MSH6 (clone 44, Ventana, prediluted), and PMS2 (clone EPR3947, Ventana, prediluted) were applied.

Hu H., Wu Z., Wang C., Huang Y., Zhang J., Cai Y., Xie X., Li J., Shen C., Li W., Ling J., Xu X, & Deng Y. (2020). Duration of FOLFOX Adjuvant Chemotherapy in High-Risk Stage II and Stage III Colon Cancer With Deficient Mismatch Repair. Frontiers in Oncology, 10, 579478.

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Immunohistochemical Detection of DNA Mismatch Repair Proteins

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Immunohistochemistry for MSH2 (clone 25D12, Novocastra (Biosystems Switzerland, Muttenz, Switzerland), dilution 1 : 100; or Clone G219-1129, Roche Ventana Medical Systems, ready-to-use), MSH-6 (clone 44, Becton Dickinson, Allschwil, Switzerland, dilution 1 : 500; or Clone EP49, Epitomics (LabForce, Nunningen, Switzerland), dilution 1 : 50), MLH1 (clone G168-15, Becton Dickinson, dilution 1 : 100; or clone M1, Roche Ventana Medical Systems, ready-to-use) and PMS2 (clone A16-4, Becton Dickinson, dilution 1 : 300; or Clone EPR3947, Roche Ventana Medical Systems, ready-to-use) was performed using the Ventana Benchmark system or using the OptiView DAB Detection Kit (Ventana Medical Systems) followed by counterstaining with haematoxylin.
Any nuclear staining in the tumour cells was considered as ‘positive'. Complete absence of immunoreactivity in the presence of a positive internal control (lymphocytes) was scored as ‘negative'.

Feilchenfeldt J., Varga Z., Siano M., Grabsch H.I., Held U., Schuknecht B., Trip A., Hamaguchi T., Gut P., Balague O., Khanfir K., Diebold J., Jochum W., Shoji H., Kushima R., Wagner D., Shimada Y., Cats A., Knuth A., Moch H., Aebi S, & Hofer S. (2015). Brain metastases in gastro-oesophageal adenocarcinoma: insights into the role of the human epidermal growth factor receptor 2 (HER2). British Journal of Cancer, 113(5), 716-721.

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Immunohistochemical Analysis of MMR Proteins

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Concerning immunohistochemistry, 4 µm-thick tissue sections were stained using a Ventana BenchMark ULTRA autostainer (Ventana Medical Systems, Tucson, Arizona) using monoclonal antibodies directed against CK7 (clone OV-TL, BioSB, 1:500), CK20 (clone KS20.8, BioSB, 1:200), MSH2 (clone G219-1129, Roche, ready to use), PMS2 (clone A16-4, Roche, ready to use), MSH6 (clone 44, Roche, ready to use), and MLH1 (clone M1, Roche, ready to use). The reactions were visualized using the Ultraview Detection System (Ventana Medical Systems). The slides were counterstained with hematoxylin, dehydrated, and then covered in a xylene-based mounting medium.

Hrudka J., Fišerová H., Jelínková K., Matěj R, & Waldauf P. (2021). Cytokeratin 7 expression as a predictor of an unfavorable prognosis in colorectal carcinoma. Scientific Reports, 11, 17863.

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Immunohistochemical Profiling of Cancer Biomarkers

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Immunohistochemistry (IHC) was performed using a Ventana XT automated stainer (Ventana) with antibodies for MutL homolog 1 (MLH1, ready to use, clone M1, Roche, Basel, Switzerland), MutS protein homolog 2 (MSH2, ready to use, clone G219-1129; Roche), MutS homolog 6 (MSH6, 1:100, clone 44; Cell Marque, Rocklin, CA, USA), postmeiotic segregation increased 2 (PMS2, 1:40, clone MRQ28; Cell Marque), epidermal growth factor receptor 2 (HER2, ready to use, clone 4B5; Roche), MET (ready to use, clone SP44; Roche), epidermal growth factor receptor (EGFR, 1:100, clone EP38Y; Abcam, Cambridge, UK), phosphatase and tensin homolog (PTEN, 1:100, clone 138G6; Cell Signaling, Danvers, MA, USA), and p53 (1:300, clone DO7; Novocastra, Newcastle, UK). IHC was performed in all cases as previously described [14] .

Woo H.Y., Bae Y.S., Kim J.H., Lee S.K., Lee Y.C., Cheong J.H., Noh S.H, & Kim H. (2017). Distinct expression profile of key molecules in crawling-type early gastric carcinoma. Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association, 20(4).

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Immunohistochemical Staining Protocol

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IHC was performed with a Ventana XT automated staining instrument. Antibodies recognizing the following targets were used: MutL homolog 1 (MLH1, ready to use, clone M1, Roche, Basel, Schweiz), MutS protein homolog 2 (MSH2, ready to use, clone G219-1129, Roche), MutS homolog 6 (MSH6, 1:100, clone 44, Cell Marque, Rocklin, CA), postmeiotic segregation increased 2 (PMS2, 1:40, clone MRQ28, Cell Marque), HER2 (ready to use, clone 4B5, Roche), EGFR (1:100, EP38Y, Abcam, Cambridge, UK), c-MET (ready to use, clone SP44, Roche), PTEN (1:100, clone 138G6, Cell signaling, Danvers, MA), and p53 (1:300, DO7, Novocastra, Newcastle, UK). Sections were deparaffinized using EZ Prep solution (Ventana Corporation, Tucson, AZ). CC1 standard (pH 8.4 buffer containing Tris/borate/EDTA) was used for antigen retrieval and blocked with inhibitor D (3% H₂O₂) for 4 min at 37°C. Slides were incubated with primary antibody for 40 min at 37°C, followed by a universal secondary antibody for 20 min at 37°C. Slides were incubated in streptavidin-horseradish peroxidase (SA-HRP) D for 16 min at 37°C, after which the substrate, 3,3′-diaminobenzidine tetrahydrochloride (DAB) H₂O₂, was added for 8 min, followed by hematoxylin and bluing reagent counterstaining at 37°C.

Kim H.S., Shin S.J., Beom S.H., Jung M., Choi Y.Y., Son T., Kim H.I., Cheong J.H., Hyung W.J., Noh S.H., Chung H., Park J.C., Shin S.K., Lee S.K., Lee Y.C., Koom W.S., Lim J.S., Chung H.C., Rha S.Y, & Kim H. (2016). Comprehensive expression profiles of gastric cancer molecular subtypes by immunohistochemistry: implications for individualized therapy. Oncotarget, 7(28), 44608-44620.

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