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Ewsr1 breakapart probe

Manufactured by Oxford Gene Technology
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The EWSR1 Breakapart probe is a molecular diagnostic tool designed to detect chromosomal rearrangements involving the EWSR1 gene. It is a fluorescence in situ hybridization (FISH) probe that can be used to identify the presence of EWSR1 gene translocations, which are commonly associated with certain types of sarcomas and other cancers.

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Lab products found in correlation

Cytovision computer assisted karyotyping system, by Leica (1 mentions) Wright s stain, by Merck Group (1 mentions) Collagenase 2, by Worthington (1 mentions) Axio scope a1, by Zeiss (1 mentions) Methanol, by Merck Group (1 mentions) Glacial acetic acid, by Merck Group (1 mentions) Cytovision system, by Leica (1 mentions) Axio imager z2, by Zeiss (1 mentions) Isis software, by MetaSystems (1 mentions) Cool cube 1 camera, by MetaSystems (1 mentions)

4 protocols using ewsr1 breakapart probe

1

Cytogenetic Analysis of Tumor Samples

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Fresh tissue from a representative area of the tumor was received and analyzed cytogenetically as part of our diagnostic routine. The samples were disaggregated mechanically and enzymatically with collagenase II (Worthington, Freehold, NJ, USA). The resulting cells were cultured and harvested using standard techniques. Chromosome preparations were G-banded with Wright's stain (SigmaAldrich; St Louis, MO, USA) and examined. Metaphases were analyzed and karyograms prepared using the CytoVision computer assisted karyotyping system (Leica Biosystems, Newcastle, UK). The karyotypes were described according to the International System for Human Cytogenetics Nomenclature [33 ].
For case 12 (see below, Table 2), FISH was performed on metaphase spreads using painting probes for chromosomes 19 and 22 as well as an EWSR1 breakapart probe (Cytocell, Cambridge, UK).
Panagopoulos I., Gorunova L., Lobmaier I., Bjerkehagen B, & Heim S. (2017). Karyotyping and analysis of GNAS locus in intramuscular myxomas. Oncotarget, 8(13), 22086-22094.
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2

Cytogenetic Analysis of Ewing's Sarcoma

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Cells were seeded on Knittel Microscope slides and cultured until 80% confluency was reached. Colchicine (1 µg/ml) was added and incubated for 2 h. Then, the cells were harvested by treatment with 0.05 M KCl (cat. no. 7477-40-7; Sigma-Aldrich; Merck KGaA) followed by fixation with 3:1 methanol (cat. no. 67-56-1; Sigma-Aldrich; Merck KGaA) and glacial acetic acid (cat. no. 100063; Merck KGaA). Finally, the slides were evaluated using the G-banding protocol (20 (link)). A total of 30 metaphases were evaluated for each patient and each cell line. Molecular cytogenetic analysis using FISH was performed to evaluate the presence of the translocation between EWSR1 (ES region 1) and any of the ETS transcription factor family genes, in particular FLI1, located in chromosomes 22 and 11, respectively. The Cytocell Aquarius® kit was used, containing the EWSR1 Breakapart Probe (Cytocell, Ltd.), with a 392-kb red probe and a 631-kb green probe to be placed on either side of the EWSR1 gene. The Carl Zeiss fluorescence microscope (Axio-Scope A1) with image capture software was used for the evaluation. The presence of fluorescent signals from the different probes in both metaphase (25 metaphase) and nucleus (200 nucleus) chromosomes were analyzed, and the findings were described according to the International Cytogenetic Nomenclature System (21 (link)).
Montoya C., Rey L., Rodríguez J., Fernández M.J., Troncoso D., Cañas A., Moreno O., Henríquez B, & Rojas A. (2020). Epigenetic control of the EWS-FLI1 promoter in Ewing's sarcoma. Oncology Reports, 43(4), 1199-1207.
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3

Chromosomal Translocations Assessment

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FISH was performed on metaphase spreads using whole painting probes for chromosomes 9 and 22, a BCR/ABL(ABL1) Translocation Dual Fusion probe (Cytocell, Cambridge, UK), and an EWSR1 Breakapart probe (Cytocell, Cambridge, UK). Fluorescent signals were captured and analyzed using the CytoVision system (Leica Biosystems, Newcastle, UK).
Panagopoulos I., Gorunova L., Bjerkehagen B, & Heim S. (2015). Fusion of the Genes EWSR1 and PBX3 in Retroperitoneal Leiomyoma with t(9;22)(q33;q12). PLoS ONE, 10(4), e0124288.
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4

FISH-based Amplification and Breakapart Analysis

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Fluorescence in-situ hybridization (FISH) was performed according to the protocol of the manufacturer using the MDM2 Amplification probe and the EWSR1 Breakapart probe (Cytocell, Cambridge, UK). Briefly, cytospins with patient cells and 2 μl probes were co-denatured at 75 °C and hybridized overnight at 37 °C. After two post-hybridization washes, fluorescent signals were viewed and analyzed with an Axio Imager Z.2 fluorescence microscope equipped with filters for DAPI, FITC, TRITC, and DEAC (Carl Zeiss AG, Feldbach, Switzerland). Images were captured with a Cool Cube 1 camera and the ISIS software (MetaSystems GmbH, Altlussheim, Germany). Two-hundred nuclei were analyzed for each hybridization.
Muff R., Botter S.M., Husmann K., Tchinda J., Selvam P., Seeli-Maduz F, & Fuchs B. (2016). Explant culture of sarcoma patients' tissue. Laboratory investigation; a journal of technical methods and pathology, 96(7).
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