Zytodot 2c cish implementation kit

Manufactured by ZytoVision

Sourced in Germany

The ZytoDot 2C CISH Implementation Kit is a laboratory tool designed for the detection of chromosome enumeration and gene amplification in cell samples using chromogenic in situ hybridization (CISH) technology. The kit provides the necessary reagents and protocols to perform CISH analysis.

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Lab products found in correlation

Zytodot 2c spec met cen7 probe, by ZytoVision (2 mentions) Clone 44, by BD (1 mentions) Bondmax detection system, by Leica (1 mentions) Eber probe, by Leica (1 mentions) Clone g168 15, by BD (1 mentions) Clone mrq28, by Cell Marque (1 mentions)

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ZytoDot CISH Implementation kit (2 citations)

5 protocols using zytodot 2c cish implementation kit

CISH-Based MET Amplification Analysis

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Analysis of MET amplification was performed by CISH using the ZytoDot 2C SPEC MET/CEN7 Probe and the ZytoDot 2C CISH Implementation Kit (ZytoVision GmbH, Bremerhaven, Germany).
The results of CISH were evaluated by screening the entire tissue sections in order to find, where present, MET‐amplified invasive cancer areas. Subsequently, MET and centromer 7 signals were counted in at least 20 representative adjacent cancer cell nuclei within the invasive region. Forty nuclei were counted when the MET/centromer 7 ratio ranged from 1.8 to 2.2. The presence of CISH clusters was noted and the ratio of MET/centromer 7 signals was calculated. The gene count was calculated by dividing the number of MET gene signals by the number of cancer cell nuclei studied.

Metzger M., Behrens H., Böger C., Haag J., Krüger S, & Röcken C. (2015). MET in gastric cancer – discarding a 10% cutoff rule. Histopathology, 68(2), 241-253.

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Comprehensive Tumor Biomarker Profiling

Cited 5 times

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The HER2-and MET-status was assessed as previously described [11] (link), [12] (link), [16] (link), [17] (link), [18] (link) using immunohistochemistry [anti-Her2/neu antibody; clone SP3, Thermo Fisher Scientific; Fremont; USA (#MA5-14509); anti-MET antibody; clone SP44; Spring Bioscience; Pleasanton, California, USA (#M3444)] and in situ–hybridization [ZytoDot 2C SPEC HER2/CEN17 Probe (#C-3032-400), ZytoDot 2C SPEC MET/CEN7 Probe (#C-3057-400) and the ZytoDot 2C CISH Implementation Kit (#C-3044-40); ZytoVision GmbH, Bremerhaven, Germany)]. Epstein–Barr virus (EBV)–encoded RNA was detected using the EBER-probe (Novocastra, Leica Microsystems GmbH, Nussloch, Germany; #PB0589) and the BondMax-detection system according to the manufacturer's instructions (Leica Microsystems GmbH). MSI status was assessed by immunostaining using antibodies directed against MLH1 (clone G168-15, BD Biosciences, Heidelberg, Germany; #MA1-25669), PMS2 (clone MRQ-28; Cell Marque Corporation, Rocklin, USA; #288M-16-ASR), MSH2 (clone FE11; Calbiochem, Merck KGaA, Darmstadt, Germany; #MABE284), and MSH6 (clone 44, BD Biosciences; #610919) as well as by comparison of the allelic profiles of the mononucleotide repeat markers BAT-25, BAT-26, NR-21, NR-24, and NR-27 in tumor and corresponding normal tissue in cases with ambiguous immunostaining [16] (link).

Schoop H., Bregenzer A., Halske C., Behrens H.M., Krüger S., Egberts J.H, & Röcken C. (2019). Therapy Resistance in Neoadjuvantly Treated Gastric Cancer and Cancer of the Gastroesophageal Junction is Associated with an Increased Expression of Immune Checkpoint Inhibitors—Comparison Against a Therapy Naïve Cohort. Translational Oncology, 13(2), 165-176.

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CISH Analysis of DLX4, HOXB3, MSX2, PTX3

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Pretreatment and denaturation of the sections was done following standard laboratory protocol using the ZytoDot 2C CISH Implementation Kit (ZytoVision GmbH, Bremerhaven, Germany). CISH probes of DLX4, HOXB3, MSX2, and PTX3 genes were used in the present study (Empire Genomics, Williamsville, NY, USA). Hybridization was done using 10 µL of each probe followed by covering of the slides using coverslip. The slides were then placed on a hot plate for 5 min at 79 °C, and transferred to a humidity chamber overnight at 37 °C. On the following day, the coverslips were removed, and DAB solution was applied to the slide per the manufacturer’s protocol. The nucleus was counterstained with a nuclear dye and dehydration using increasing gradients of ethanol was done. Cover slip was placed, and the hybridized probe fragments were visualized using brightfield microscope. Under the microscope, two brown coloured dots (signals) were expected per nuclei of normal cells in interphase or metaphase without aberrations of the examined chromosomes. Due to limited availability of tissue material, only three control samples were used for CISH (1 female child and 2 male children).

Jain N, & Pilmane M. (2021). Evaluating the Expression of Candidate Homeobox Genes and Their Role in Local-Site Inflammation in Mucosal Tissue Obtained from Children with Non-Syndromic Cleft Lip and Palate. Journal of Personalized Medicine, 11(11), 1135.

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CISH Protocol for 1p and 19q Assessment

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CISH of 1p and 19q was performed by ZytoDot 2C CISH Implementation Kit (ZytoVision, Bremerhaven, Germany). After deparaffinization and blocking, pretreatment was performed with EDTA Pretreatment Solution at 95°C. After incubation with pepsine samples were rehydrated. 10μl of probes were added, fixed with Fixogum and denatured at 79°C. Hybridization was performed over night at 37°C. Next, slides were incubated with Anti-DIG and HRP/AP-Polymer. The 1p or 19q probe was visualized by AP Red Solution, the control probe by HRP-Green Solution. Slides were counterstained with Nuclear Blue followed by washing, dehydration and mounting. Loss of heterozygosity (LOH) was determined according to the guidelines of the Research Committee of the European Confederation of Neuropathological Societies [36 (link)].

Mäder L., Blank A.E., Capper D., Jansong J., Baumgarten P., Wirsik N.M., Zachskorn C., Ehlers J., Seifert M., Klink B., Liebner S., Niclou S., Naumann U., Harter P.N, & Mittelbronn M. (2018). Pericytes/vessel-associated mural cells (VAMCs) are the major source of key epithelial-mesenchymal transition (EMT) factors SLUG and TWIST in human glioma. Oncotarget, 9(35), 24041-24053.

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CISH Analysis of FGFR2 Amplification

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Analysis of Fgfr2 amplification was analyzed by CISH using the ZytoDot^® 2C (SPEC Fgfr2/CEN 10 Probe) and the ZytoDot 2C CISH Implementation Kit (ZytoVision GmbH, Bremerhaven, Germany). Fgfr2-CISH was evaluated by screening the entire tissue section to find Fgfr2 amplified tumor cells. Fgfr2 and centromere 10 signals were counted in at least 20 representative adjacent tumor cell nuclei within the invasive regions. The Fgfr2/centromere ratio of 10 was then calculated. Probes with a ratio greater than 2.2 were classified as Fgfr2 amplified. Probes with a ratio lower 1.8 were classified as non-amplified. Forty nuclei were counted if the Fgfr2/centromere 10 ratio ranged from 1.8 to 2.2. If the ratio was still in the range of 1.8 to 2.2, a cutoff of >2.0 was used to classify probes as Fgfr2 amplified. CISH clusters were also observed. Tumors with an average Fgfr2 count of greater than 4 signals per nucleus were classified as Fgfr2 polysomic.

Schrumpf T., Behrens H.M., Haag J., Krüger S, & Röcken C. (2022). FGFR2 overexpression and compromised survival in diffuse-type gastric cancer in a large central European cohort. PLoS ONE, 17(2), e0264011.

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