A16 4

Manufactured by Roche

Sourced in United States

The A16-4 is a high-performance laboratory instrument designed for a variety of analytical applications. It features precise temperature control, advanced data processing capabilities, and a user-friendly interface. The core function of the A16-4 is to provide reliable and accurate measurements for research and diagnostic purposes.

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Lab products found in correlation

G219 1129, by Roche (4 mentions) Optiview dab detection kit, by Roche (1 mentions) Optiview amplification kit, by Roche (1 mentions) Ventana benchmark ultra, by Roche (1 mentions) Benchmark ultra, by Roche (1 mentions) Benchmark xt, by Roche (1 mentions)

Spelling variants of this product

PMS2 (clone A16-4, (4 citations) Clone A16‐4 (4 citations)

5 protocols using a16 4

Immunohistochemical Analysis of Mismatch Repair Proteins

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Tissues were cut into 4-μm-thick sections. The MMR panel consisted of monoclonal antibodies—mouse anti-MLH1 (Ventana, M1), mouse anti-PMS2 (Ventana, A16-4), mouse anti-MSH2 (Ventana G219-1129), and rabbit anti-MSH6 (Ventana, SP93). The panel was performed using the Ventana Optiview DAB Detection kit, the Ventana Optiview Amplification Kit, and ancillaries on the fully automated Roche Ventana Ultra instrument. MMR loss was determined when the tumor showed loss of expression for the examined MMR proteins. Normal tissue adjacent to the tumor was used as a positive internal control.

Kumar V., Ramnarayanan K., Sundar R., Padmanabhan N., Srivastava S., Koiwa M., Yasuda T., Koh V., Huang K.K., Tay S.T., Ho S.W., Tan A.L., Ishimoto T., Kim G., Shabbir A., Chen Q., Zhang B., Xu S., Lam K.P., Lum H.Y., Teh M., Yong W.P., So J.B, & Tan P. (2022). Single-Cell Atlas of Lineage States, Tumor Microenvironment, and Subtype-Specific Expression Programs in Gastric Cancer. Cancer Discovery, 12(3), 670-691.

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Immunohistochemical Evaluation of DNA-MMR Proteins

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Immunohistochemical staining of DNA-MMR protein expression was performed on endometrial and ovarian samples in blocks fixed with formalin and embedded in paraffin. Representative whole 5-μm-thick sections were performed with IHC. Anti-PMS2 (A16-4, 1:200; Ventana Medical Systems, Arizona, USA), anti-MLH1 (M1, 1:200 dilution; Ventana Medical Systems, Arizona, USA), anti-MSH2 (G219-11229, 1:50 dilution, Ventana Medical Systems, Arizona, USA), and anti-MSH6 (SP93, 1:200 dilution; Ventana Medical Systems, Arizona, USA) were evaluated. Tissues were stained with antibodies against PMS2, MSH6, MLH1, and MSH2 following deparaffinization. Slides were counterstained with hematoxylin & eosin. Complete loss of nuclear staining for at least one MMR protein was defined as MMR deficiency. Stromal and inflammatory cells of the adjacent normal mucosa with intact nuclear staining were used as an internal positive control.

Gezer Ş, & Bayrak B.Y. (2021). Metastatic and synchronous ovarian involvement in low-risk endometrial cancer; clinicopathological analysis with detection of DNA mismatch repair deficiency. Ginekologia polska.

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Assessing Mismatch Repair and BRAF Status

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Immunohistochemical analysis for MMR status was carried out on 3-μm sections prepared from TMA blocks and subjected to immunohistochemistry for the mutL homologue 1 (MLH1; G168-15, Roche Diagnostics), mutS homologue 2 (MSH2; G219-1129, Roche Diagnostics), mutS homologue 6 (MSH6; SP93, Roche Diagnostics) and postmeiotic segregation increased 1 homologue 2 (PMS2; A16-4, Roche Diagnostics) with the Ventana Benchmark Ultra (Roche Diagnostics). We defined dMMR as complete absence of nuclear staining with 1 or more of the 4 antibodies in the tumoural cells in the presence of positive internal control (nuclei of lymphocytes and stromal cells). BRAF mutation status was assessed by immunohistochemistry with anti-V600E-mutant-BRAF antibody (VE1, Roche Diagnostics). Positive results were defined as granular cytoplasmic staining of the tumoural cells.

Evaristo G., Katz A., Ramírez-GarcíaLuna J.L., Issac M.S., Sangwan V., Thai D.V., Bertos N., Guiot M.C., Camilleri-Broët S., Marcus V., Mueller C., Cools-Lartigue J., Fiset P.O, & Ferri L.E. (2023). Relation between mismatch repair status, chemoresponse, survival and anatomic location in gastroesophageal adenocarcinoma. Canadian Journal of Surgery, 66(1), E79-E87.

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Immunohistochemical Detection of dMMR Proteins

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Immunohistochemistry was performed on 4-µm paraffin sections. The primary antibodies specific for dMMR proteins were as follows: MSH2 (Roche G219-1129), MLH1 (Roche M1), MSH6 (Abcam EPR3945), and PMS2 (Roche A16-4). All dMMR protein staining was performed using an autoimmunostainer (BenchMark Ultra, Ventana Medical Systems).

Shimizu K., Sano T., Mizuno K., Sunada T., Makita N., Hagimoto H., Goto T., Sawada A., Fujimoto M., Ichioka K., Ogawa O., Kobayashi T, & Akamatsu S. (2022). A case of microsatellite instability-high clinically advanced castration-resistant prostate cancer showing a remarkable response to pembrolizumab sustained over at least 18 months. Cold Spring Harbor Molecular Case Studies, 8(4), a006194.

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Automated Immunostaining for MMR and EBV

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Fully automated immunostaining was performed on 4-μm-thick FFPE sections using a BenchMark XT autostainer (Ventana Medical Systems, Tucson, AZ). IHC for MMR proteins was performed according to standard antibody protocols using the following antibodies: mouse anti-MLH1 (Ventana/Roche M1, pre-dilute antibody), mouse anti-MSH2 (Roche G219-1129, pre-dilute antibody), rabbit anti-MSH6 (Roche SP93, pre-dilute antibody), and mouse anti-PMS2 (Roche A16-4, pre-dilute antibody). Tumors lacking MLH1, MSH2, PMS2, or MSH6 expression were considered dMMR, whereas tumors that expressed MLH1, MSH2, PMS2, or MSH6 were considered pMMR.
EBV status was determined by in situ hybridization with EBV-encoded small RNA (EBER) probes (INFORM EBER, Roche, ready-to-use) according to product protocols. Strong EBER signals were interpreted as EBV-positive.

Son S.M., Woo C.G., Kim D.H., Yun H.Y., Kim H., Kim H.K., Yang Y., Kwon J., Kwon M., Kim T.Y., Kim H.D., Koh J.Y., Park S.H., Shin E.C, & Han H.S. (2020). Distinct tumor immune microenvironments in primary and metastatic lesions in gastric cancer patients. Scientific Reports, 10, 14293.

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