Fh535

Manufactured by Selleck Chemicals

Sourced in United States

The FH535 is a laboratory centrifuge designed for rapid separation of liquids and solids. It features a fixed-angle rotor that can accommodate a range of sample tube sizes. The centrifuge operates at a maximum speed of 5,350 rpm and provides a maximum relative centrifugal force of 3,535 x g.

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Lab products found in correlation

8 protocols using fh535

Comprehensive Autophagy and Signaling Pathway Analysis

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The antibodies used in our experiments were: LC3 (CST, #2775), IL-33 (R & D, AF3626 and AF4810), ST2 (Thermofisher, PA5-20077), p-STAT3 (CST, #9145), STAT3 (CST, #9139), p-AMPK (CST, #2535), AMPK (CST, #2532), p-ULK1 (Thermofisher, PA5-105129), ULK1 (Abclonal, A8529), p62 (Abclonal, A1970), GKN-1 (Proteintech, 19344-1-AP), GKN-2 (Abcam, ab188866), GKN-3 (Cloud-clone, PAK528Mu01), Tubulin (Proteintech, 66031-1-Ig).
The chemicals used in our experiments were: Bafilomycin A1 (Selleck, S1413), 3-MA (Selleck, S2767), Rapamycin (Selleck, S1039), MNNG (Selleck, E0157), NAC (Selleck, S1623), Cycloheximide (Selleck, S7418), DAPI (Beyotime, C1002), Recombinant Human IL-33 (R & D, 3625-IL), Pifithrin-α HBr (Selleck, E0157), 2-MeOE2 (Selleck, S1233), ML385 (Selleck, S8790), SR11302 (MCE, HY-15870), BAY 11 (Selleck, S2913), GW9662 (Selleck, S2915), IQ3 (Selleck, S0781), STAT3-IN-7 (Selleck, S0986), FH535 (Selleck, S7484).

Liu K., Huang H., Xiong M., Wang Q., Chen X., Feng Y., Ma H., Chen W., Li X, & Ye X. (2024). IL-33 Accelerates Chronic Atrophic Gastritis through AMPK-ULK1 Axis Mediated Autolysosomal Degradation of GKN1. International Journal of Biological Sciences, 20(6), 2323-2338.

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Investigating Wnt/β-Catenin Signaling

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FH535 was purchased from Selleckchem (S7484) and soluted in DMSO (Sigma-Aldrich). In all in vitro assays, DMSO was used as control, at final concentrations no more than 0.1%. Primary antibodies against β-catenin, Cyclin D1, survivin, Snail, vimentin and β-actin for Western blotting were obtained from Cell Signaling Technology.

Chen Y., Rao X., Huang K., Jiang X., Wang H, & Teng L. (2017). FH535 Inhibits Proliferation and Motility of Colon Cancer Cells by Targeting Wnt/β-catenin Signaling Pathway. Journal of Cancer, 8(16), 3142-3153.

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FH535 Compound Preparation Protocol

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FH535 was purchased from Selleck, and dissolved in DMSO at a concentration of 50 mmol/L as a stock solution and used by diluting in DMSO to give a final concentration of 50 μmol/L 17.

Wen D., Liao T., Ma B., Qu N., Shi R., Lu Z., Wang Y., Wei W, & Ji Q. (2018). Downregulation of CSN6 attenuates papillary thyroid carcinoma progression by reducing Wnt/β‐catenin signaling and sensitizes cancer cells to FH535 therapy. Cancer Medicine, 7(2), 285-296.

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Kinase Inhibitor Assay Protocol

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Imatinib, nilotinib, dasatinib, bafetinib, ponatinib, DCC-2036, GNF-2, GNF-5, EPZ005687, XAV-939, and FH535 were purchased from Selleckchem (Houston, TX, USA). PHA665752 was purchased from MedChemExpress, LLC (Princeton, NJ, USA). U0126, LY294002, and SP600125 were purchased form Wako (Tokyo, Japan). These reagents were dissolved in dimethyl sulfoxide and diluted in phosphate-buffed saline (PBS; 0.05 M, pH 7.4), filtrated through syringe filters (0.45 μm, IWAKI GLASS, Tokyo, Japan), and used for various assays described below.

Tsubaki M., Takeda T., Kino T., Sakai K., Itoh T., Imano M., Nakayama T., Nishio K., Satou T, & Nishida S. (2017). Contributions of MET activation to BCR-ABL1 tyrosine kinase inhibitor resistance in chronic myeloid leukemia cells. Oncotarget, 8(24), 38717-38730.

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Evaluating Small Molecule Inhibitors on CCOs

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CCOs were cultured in 24-well plates for 2 weeks, and then harvested and dissociated using TrypLE Express. The dissociated CCOs were mixed with MBM + Matrigel (1:3 ratio) and seeded in 96-well white plates (10 μL of 2 × 10³ cells/well; Corning). After gelation, 100 μL MBM was added to each well. The CCOs were allowed to grow for 10–14 days until their diameters reached 100 μm. Then, ten concentrations of FH535, ICG-001 (all Selleckchem, TX, USA), and DMSO controls were added every 3 days. After 6 days, the medium was changed to 100 μL MBM per well to measure cell viability, and 100 μL CellTiter-Glo (Promega) was added to each well. The experiments were conducted in triplicate. The plates were agitated for 30 min at room temperature prior to luminescence measurement. IC₅₀ values were determined using Graph Pad Prism (v5.01).

Cho E.J., Kim M., Jo D., Kim J., Oh J.H., Chung H.C., Lee S.H., Kim D., Chun S.M., Kim J., Lee H., Kim T.W., Yu C.S., Sung C.O, & Jang S.J. (2021). Immuno-genomic classification of colorectal cancer organoids reveals cancer cells with intrinsic immunogenic properties associated with patient survival. Journal of Experimental & Clinical Cancer Research : CR, 40, 230.

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Hedgehog Signaling Modulation in Vitro and In Vivo

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For in vitro experiments, the IWP2 (Selleck Chemicals, USA) and FH535 (Selleck Chemicals, USA) were dissolved in dimethyl sulfoxide and applied at a final concentration of 10 μM and 20 μM in cell culture medium, respectively. Recombinant human sonic hedgehog (R and D systems, USA) was dissolved in PBS and applied at a final concentration of 5 μg/ml in cell culture medium.
For in vivo experiments, Cyclopamine (Selleck Chemicals, USA), GANT61 (Selleck Chemicals, USA), and IWP2 were reconstituted in corn oil. Mice were treated with 20 mg/kg Cyclopamine, 30 mg/kg GANT61, or 10 mg/kg IWP2 through intragastric administration every other day after tamoxifen injection. Mice were treated for 2 months and then evaluated.

Deng Q., Li P., Che M., Liu J., Biswas S., Ma G., He L., Wei Z., Zhang Z., Yang Y., Liu H, & Li B. (2019). Activation of hedgehog signaling in mesenchymal stem cells induces cartilage and bone tumor formation via Wnt/β-Catenin. eLife, 8, e50208.

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Cell Proliferation and Drug Sensitivity Assay

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Following transfection, cell proliferation was assessed using CCK8 assay (DOJINDO, Japan), according to the manufacturer's instructions. Forty-eight hours after transfection with 20 nM miR-506-3p, or EZH2, or 50uM si-EZH2, or β-catenin or coculture with 30uM β-catenin inhibitor FH535 (Selleck, USA), 1.3 × 10³ cells per well were seeded in 96-well plates and treated with a titration of cisplatin (DOJINDO, Japan) or olaparib (Selleck, USA). The medium and drug were supplemented at day 3 for cisplatin or olaparib treatment. After incubation for 5 (cisplatin) or 7 (olaparib) days, cell viability was estimated and surviving fractions were calculated. Cell survival was calculated by normalizing the absorbance to that of untreated controls.

Sun Y., Wu J., Dong X., Zhang J., Meng C, & Liu G. (2020). MicroRNA-506-3p increases the response to PARP inhibitors and cisplatin by targeting EZH2/β-catenin in serous ovarian cancers. Translational Oncology, 14(2), 100987.

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Inflammatory Signaling Pathway Modulation

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We used the following reagents: lipopolysaccharide (LPS; L4391, Sigma-Aldrich, St. Louis, MO); IL-6 (RMIL6I, Thermo Fisher Scientific, Waltham, MA), IL-1β (I5271, Sigma-Aldrich St. Louis, MO); TNF-α (RMTNFAI, Thermo Fisher Scientific, Waltham, MA); forskolin (#1099, TOCRIS, Bristol, United Kingdom); LY2090314 (SML1438, Sigma-Aldrich); CHIR99021 (SML1046, Sigma-Aldrich St. Louis, MO); methyl 3- 4methylphenyl sulfonyl amino benzoate (MSAB; M60316, Xcess Biosciences, San Diego, CA); and FH535 (S7484, SelleckChem, Houston, TX).

Cordonier E.L., Liu T., Saito K., Chen S.S., Xu Y, & Fukuda M. (2019). Luciferase Reporter Mice for In Vivo Monitoring and Ex Vivo Assessment of Hypothalamic Signaling of Socs3 Expression. Journal of the Endocrine Society, 3(7), 1246-1260.

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