Protease solution

Manufactured by Abbott

Sourced in United States

Protease solution is a laboratory reagent used for the digestion and breakdown of protein molecules. It contains a mixture of enzymes that catalyze the hydrolysis of peptide bonds, allowing for the separation and isolation of proteins from complex samples.

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Lab products found in correlation

Pretreatment solution, by Abbott (3 mentions) Hybrite, by Abbott (1 mentions) Pathvysion assay, by Abbott (1 mentions) Spectrum orange, by Abbott (1 mentions) Nick translation kit, by Abbott (1 mentions) Large construction kit, by Qiagen (1 mentions)

4 protocols using protease solution

Fluorescence In-Situ Hybridization for HER2 and CIN

Cited 2 times

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To identify the HER2 status and CEP17 copy number in each case, HER2 FISH (PathVysion assay, Abbott Molecular, Downers Grove, IL) was performed on TMAs of the first set and all tissue sections of the second set. FISH using CEP1 [Vysis CEP1 (D1Z5) SpectrumOrange Probe, Abbott Molecular], CEP8 [Vysis CEP8 (D8Z2) SpectrumGreen Probe, Abbott Molecular], CEP11 [Vysis CEP11 (D11Z1) SpectrumGreen Probe, Abbott Molecular], and CEP16 probe [Vysis CEP16 (D16Z3) SpectrumGreen Probe, Abbott Molecular] was performed on TMAs to assess CIN. These CEP probes around the centromere have been reported to show frequent copy number gains in breast cancer^{17 (link),23 (link),28 (link)}.
Briefly, 4 μm deparaffinized tissue sections were incubated in pretreatment solution (Abbott Molecular) at 80 °C for 30 min and then, in protease solution (Abbott Molecular) at 37 °C for 20 min. Probes were diluted in tDen-Hyb-2 hybridization buffer (Insitus Biotechnologies, Albuquerque, NM). The probes and the DNA from the tissue sections were denatured together by incubating them for 5 min at 73 °C in HYBrite™ (Abbott Molecular), and then hybridized for 16 h at 37 °C. Post-hybridization washes were performed according to the manufacturer’s protocol. The mounted slides were viewed using a fluorescence microscope.

Lee K., Kim H.J., Jang M.H., Lee S., Ahn S, & Park S.Y. (2019). Centromere 17 copy number gain reflects chromosomal instability in breast cancer. Scientific Reports, 9, 17968.

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Fluorescent In Situ Hybridization for Y Chromosome Detection

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Four patients had a sex mismatch with donor lung grafts. We confirmed the tumor origin results evidenced by STR using fluorescent in situ hybridization (FISH) targeting the sexual Y chromosome. The Y chromosome in the tumor and surrounding normal lung graft tissue was examined. FISH for Chromosome Y gene detection was performed as follows: First, 3 µm-thick FFPE tissue slides were dewaxed, dehydrated, cooked for 12 min in a pretreatment buffer at 80°C (Abbott Molecular, IL, USA), and then treated with protease solution (Abbott Molecular, IL, USA) for 14 min at 37°C. Hybridization was then performed with SureFISH ChrY CEP 273kb RD (Agilent Technologies, Santa Clara, California, USA), at 82°C for 5 min and then at 37°C for 22 h in a ThermoBrite Statspin Hybridizer (Abbott Molecular, IL, USA). Non-specific binding was removed by immerging the slides in a wash buffer I (0.7XSSC, 0.3% NP40 pH 7) 10 min at room temperature then in the wash buffer II (2XSSC, 0.1% NP40 pH 7) at 65°C 5 min. The slides were then rinsed three times in distilled water, dehydrated, air-dried, and mounted with VECTASHIELD Antifade Mounting Medium with DAPI (Clinisciences, Nanterre, France) and a cover glass. The samples were stored at 4°C until further use.

De Wolf J., Robin E., Vallee A., Cohen J., Hamid A., Roux A., Leguen M., Beaurepere R., Bieche I., Masliah-Planchon J., Glorion M., Allory Y, & Sage E. (2023). Donor/recipient origin of lung cancer after lung transplantation by DNA short tandem repeat analysis. Frontiers in Oncology, 13, 1225538.

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Fluorescence In Situ Hybridization of FFPE Tissue

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Formalin-fixed paraffin-embedded tissue samples were cut to a thickness of 5 μm and mounted on positively charged slides. Slides were heated at 60°C for 40 minutes, then deparaffinized by washing in Citrisolve 3 times for 5 minutes each, followed by two washes in 100% ethanol for 1 minute each. Slides were air dried, then immersed in Pretreatment solution (Abbott Molecular, Abbott Park, IL, USA) at 85°C for 20 minutes, rinsed in distilled water for 3 minutes, then immersed in Protease solution (Abbott Molecular, Abbott Park, IL, USA) at 37°C for 20 minutes. Slides were rinsed in distilled water for 3 minutes then air dried, followed by immersion in 70% ethanol, 85% ethanol and 100% ethanol for 1 minute each, then air dried again. Vysis CEP X SpectrumOrange/CEP Y SpectrumGreen direct labeled fluorescent DNA probe (Abbott Molecular, Abbott Park, IL, USA) was applied to the tissue, which was then coverslipped and codenatured by heating at 73°C for 5 minutes. Slides were hybridized overnight at 37°C in a humidified chamber. Slides were washed in 2xSSC/0.3% NP-40 at 74°C for 2 minutes, then 2xSSC/0.3% NP-40 at room temperature for 1 minute. Slides were counterstained with Vysis DAPI II Counter-stain (Abbott Molecular, Abbott Park, IL, USA).

Broestl L., Rubin J.B, & Dahiya S. (2017). Fetal microchimerism in human brain tumors. Brain Pathology, 28(4), 484-494.

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FGFR1 and FGFR2 FISH Analysis

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Four-micrometer deparaffinised section was incubated in pre-treatment solution (Abbott Molecular, Downers Grove, IL) at 80 °C for 40 min and then in protease solution (Abbott Molecular) for 40 min at 37 °C. Fibroblast growth factor receptor 1 (FGFR1) FISH was performed with locus-specific bacterial artificial chromosome (BAC), RP11-100B16 (chr8:38,358,839-38,522,417). We obtained the BAC clone from Invitrogen (Carlsbad, CA) and purified it with a large construction kit (Qiagen, Valencia, CA). DNA from the BAC clone was labelled with SpectrumOrange using a nick translation kit (Abbott Molecular). FGFR1 BAC probe and FGFR2 probe (Vysis LSI FGFR2 SpectrumOrange Probe, 08N42-020, Abbott Molecular) were denatured at 73 °C for 5 min and hybridised at 37 °C for 20 h. Post-hybridisation washes were performed according to the protocol supplemented. Slides were mounted in 4′,6-diamidino-2-phenylindole/anti-fade and viewed with a fluorescence microscope. The FGFR1 or FGFR2 was considered to be amplified if the average gene copy number was ≥6, and high-level amplification was defined as an average gene copy number ≥10.

Ahn S., Kim H.J., Kang E., Kim E.K., Kim S.H., Kim J.H., Kim I.A, & Park S.Y. (2020). Genomic profiling of multiple breast cancer reveals inter-lesional heterogeneity. British Journal of Cancer, 122(5), 697-704.

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