Chameleon plate reader

Manufactured by Hidex

Sourced in Finland, Austria

The Chameleon plate reader is a high-performance spectrophotometer designed for a wide range of applications in life science research. It features a compact and ergonomic design, with a touchscreen interface for easy operation. The Chameleon plate reader is capable of absorbance, fluorescence, and luminescence measurements across multiple wavelengths, allowing for versatile and accurate data collection.

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Lab products found in correlation

3h all trans retinol, by PerkinElmer (2 mentions) Rpnq0006, by PerkinElmer (2 mentions) Ez link sulfo nhs lc biotinylation kit, by Thermo Fisher Scientific (2 mentions) Luciferase assay system kit, by Promega (2 mentions) Jetprime reagent, by Polyplus Transfection (2 mentions) Celltiter glo 3d cell viability assay, by Promega (2 mentions) Vmax microplate reader, by Molecular Devices (1 mentions) Fv1000 inverted confocal microscope, by Olympus (1 mentions) Las x1, by Leica camera (1 mentions) Dmi8 inverted microscope, by Leica camera (1 mentions) Brdu cell proliferation kit, by Cell Signaling Technology (1 mentions) Prism 7, by GraphPad (1 mentions) Jetprime, by Polyplus Transfection (1 mentions) Atp determination kit, by Thermo Fisher Scientific (1 mentions) Igg2c antibodies, by Southern Biotech (1 mentions) Hrp conjugated secondary antibody, by Agilent Technologies (1 mentions) Emax plate reader, by Molecular Devices (1 mentions) Megascript t7 kit, by Thermo Fisher Scientific (1 mentions) Megaclear kit, by Thermo Fisher Scientific (1 mentions) Transit mrna transfection kit, by Mirus Bio (1 mentions)

29 protocols using chameleon plate reader

Quantitative Metabolic Profiling of Flies

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ATP levels were assayed using the ATP Determination Kit (Life Technologies, Carlsbad, CA) according to the manufacturer’s instructions. For each sample, a set of ten flies were homogenized in 100 μl of 6 M guanidine hydrochloride and then centrifuged at 12,000 g_max for 5 minutes at 4°C. Supernatants were diluted tenfold in TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0), and 10 μl of the diluted sample was mixed with the reaction solution. Different genotypes were processed in biological triplicates and luminescence was measured using a Chameleon plate reader (Hidex, Turku, Finland).
Lactate was assayed by using a BioVision Lactate Colorimetric/Fluorometric Assay Kit (BioVision, Milpitas, CA) according to the manufacturer’s instructions. Samples of 30 flies per genotype were homogenized in 300 μl of lactate assay buffer and centrifuged at 12,000 g_max for 5 minutes at 4°C. Each genotype was analyzed in triplicate and sample fluorescence was recorded using the Hidex Chameleon plate reader.

Vartiainen S., Chen S., George J., Tuomela T., Luoto K.R., O’Dell K.M, & Jacobs H.T. (2014). Phenotypic rescue of a Drosophila model of mitochondrial ANT1 disease. Disease Models & Mechanisms, 7(6), 635-648.

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Radiometric Assay for RBP4 Binding

Cited 2 times

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Compound binding to RBP4 was assessed in the radiometric scintillation proximity (SPA) assay that was previously described.^{4 (link)–7 (link)} The assay measured competitive displacement of radiolabeled [³H]-all-trans-retinol from native RBP4 purified from human urine (Fitzgerald, 30R-AR022L). The protein was biotinylated using the EZ-link Sulfo-NHS-LC-Biotinylation kit from ThermoFisher (Cat #21335) as recommended by the manufacturer. Binding assays were implemented in a final volume of 100 μL in SPA buffer (1× PBR, pH 7.4, 1 mM ethylenediaminetetraacetic acid (EDTA), 0.1% BRA, 0.5% CHAPS). The assay reaction included a radioligand, 10 nM [³H]-all-trans-retinol (48.7 Ci/mmol; PerkinElmer, Waltham, MA), along with the 0.3 mg/well streptavidin-PVT beads (PerkinElmer, RPNQ0006) and 50 nM biotinylated human RBP4. Unlabeled retinol (Sigma-Aldrich, cat # 95144) at 20 μM was added to control wells to assess a nonspecific binding. Radioactivity counts were measured using CHAMELEON plate reader (Hidex Oy, Turku, Finland) after 16 h of incubation at rt with mild shaking.

Cioffi C.L., Raja A., Muthuraman P., Jayaraman A., Jayakumar S., Varadi A., Racz B, & Petrukhin K. (2021). Identification of Transthyretin Tetramer Kinetic Stabilizers That Are Capable of Inhibiting the Retinol-Dependent Retinol Binding Protein 4-Transthyretin Interaction: Potential Novel Therapeutics for Macular Degeneration, Transthyretin Amyloidosis, and Their Common Age-Related Comorbidities. Journal of medicinal chemistry, 64(13), 9010-9041.

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Quantifying Serum Immunoglobulins and Autoantibodies

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Serum Ig levels were determined by ELISA using sheep anti-mouse Ig antibodies (Silenus Laboratories, Hawthorn, VIC, Australia) and mouse Ig isotype-specific goat antibodies conjugated with HRP (Southern Biotech) as previously described.^{60 (link)} Absorbance was read at 405 nm using a VMax microplate reader (Molecular Devices, Sunnyvale, CA, USA). ANAs were quantitated using ELISA kits for detecting mouse anti-dsDNA IgG, anti-ssDNA IgG, anti-dsDNA IgM, anti-ssDNA IgM (Alpha Diagnostics International, San Antonio, TX, USA). Absorbance was read at 450 nm using a Chameleon plate reader (Hidex, Broomhill, UK). ANAs were also detected by confocal microscopy using the ANA Test system (Immuno Concepts, Sacramento, CA, USA) and slides coated with HEp-2 human epithelial cells, stained according to the manufacturer's instructions, as described.^{60 (link)} Images were acquired using an Olympus FV1000 inverted confocal microscope (Olympus) (60 × 1.4N.A with oil). ANA levels were assessed semiquantitatively according to the brightness of fluorescence intensity on a scale of 0 to 3+.

Anstee N.S., Vandenberg C.J., Campbell K.J., Hughes P.D., O'Reilly L.A, & Cory S. (2016). Overexpression of Mcl-1 exacerbates lymphocyte accumulation and autoimmune kidney disease in lpr mice. Cell Death and Differentiation, 24(3), 397-408.

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ATP Quantification in NALM-6 Cells

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ATP was quantified using a Luminescent ATP Detection Assay Kit (ab113849, ABCAM). Briefly, 2 × 10⁴ NALM-6 cells were seeded in triplicate in a white 96-well plate. They were treated with 2DG (1 or 10 mM), with or without mannose (10 mM), for 0.5, 1, 3, and 6 h. After cell lysis with the detergent solution provided, D-luciferin and luciferase were added, and 10 min later, luminescence was measured with a Hidex Chameleon plate reader. The light emitted during the reaction is proportional to the ATP present.

Tailler M., Lindqvist L.M., Gibson L, & Adams J.M. (2018). By reducing global mRNA translation in several ways, 2-deoxyglucose lowers MCL-1 protein and sensitizes hemopoietic tumor cells to BH3 mimetic ABT737. Cell Death and Differentiation, 26(9), 1766-1781.

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Evaluating CBX-mediated FOXO3 Inhibition

Cited 1 time

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To determine CBX-mediated inhibition of FOXO3 binding to the DEPP- and BIM-promoter, a luciferase reporter plasmid containing the DEPP-promoter [38 (link)] or the BIM-promoter [39 (link)] was transiently transfected into SH-EP/FOXO3 cells using the JetPrime^® Reagent (Polyplus, Berkeley, USA) according to the manufacturer’s instructions. Subsequently the cells were cultured in the presence of 4OHT alone, or in combination with increasing concentrations of CBX for eight hours. Luciferase activity was measured with the Luciferase Assay System kit (Promega, Madison, USA) according to the manufacturer’s instructions. The luminescence signal was measured with the chameleon plate reader (Hidex, Turku, Finland).

Salcher S., Spoden G., Hagenbuchner J., Führer S., Kaserer T., Tollinger M., Huber-Cantonati P., Gruber T., Schuster D., Gust R., Zwierzina H., Müller T., Kiechl-Kohlendorfer U., Ausserlechner M.J, & Obexer P. (2019). A drug library screen identifies Carbenoxolone as novel FOXO inhibitor that overcomes FOXO3-mediated chemoprotection in high-stage neuroblastoma. Oncogene, 39(5), 1080-1097.

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Anticancer Activity of Anastrozole

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The MTT assay was carried out on MCF7, HepG2, and PC3 to determine the anticancer activity of anastrozole. Anastrozole was dissolved in Dimethylsulfoxide (DMSO) to produce the stock solution. Doxorubicin was used as a control in each plate and incubated. After removing the media, the obtained formazan crystals were stabilized by the addition of DMSO. The absorbance was measured (570 nm) using a Hidex Chameleon plate reader and the growth inhibition was calculated. The kit used contains Hidex Chamelon plate reader with a 570 nm filter, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; 10 × 5 mg), multi-channel pipette (8 to 12 channel), pipette tips (10-100 µL), phosphate-buffered saline (PBS), sterile HCl, DMSO; 0.01 M and CO₂ incubator (5%).

Hassan F., El-Hiti G.A., Abd-Allateef M, & Yousif E. (2017). Cytotoxicity anticancer activities of anastrozole against breast, liver hepatocellular, and prostate cancer cells. Saudi Medical Journal, 38(4), 359-365.

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3D Spheroid Formation and Viability Assessment

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For the production of 3D spheroids, the GravityPLUS™ microtissue culture system (InSphero AG, Zürich, Switzerland) was used. A total of 2 × 10⁴ cells were seeded in 40 μl drops into the hanging-drop plates and grown for 96 hours to form spheroids. Size was monitored regularly by live-cell microscopy. Subsequently, the spheroids were treated with etoposide alone or in combination with CBX for another 96 hours. Viable cells were stained with 2 µM calcein-AM for two hours at 37 °C. Representative live-cell images of the spheroids were taken with the DMi8 inverted microscope (Leica, Germany) and processed with the LAS X1.1.0 software (Leica, Germany). Quantification of living cells was done using the CellTiter-Glo 3D^® cell viability assay according to the manufacturer’s instructions (Promega, Madison, USA). Briefly, single spheroids were collected in 50 µl RPMI1640 media and incubated with 50 µl CellTiter-Glo 3D^® cell viability reagent for 30 min at room temperature. The luminescence signal was measured with the chameleon plate reader (Hidex, Turku, Finland).

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BrdU Proliferation Assay in Breast Cancer Cells

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HMEC, MDA‐MB‐231 and MCF‐7 cells were seeded onto 96‐well plates (1 × 10⁴ cells/well) in respective media supplemented with 10% FBS. Cells were allowed to adhere for 3 hours at 37°C before switching to media supplemented with 2% FBS. Cells were incubated overnight in low serum (LS) conditions prior to initiating treatment. Cells were treated with respective media supplemented with 10% or 2% FBS and containing BAY11‐7082 (2.5 µmol/L), UNC1999 (1 µmol/L), Tazemetostat (5 µmol/L) or non‐treated control (0.1% DMSO). Prior to treatment, 5‐bromo‐2'‐deoxyuridine (BrdU) was added to media at a final 1X concentration. BrdU incorporation was assayed after 24 hours according to manufacturer's instructions (BrdU Cell Proliferation Kit, Cell Signaling Technology). Absorbance was measured at 450 nm using the Hidex Chameleon plate reader. Treatments were performed in quadruplicate (n = 4) and tested for statistical significance using one‐way ANOVA on Graphpad Prism 7.

Duan S., Chan W.K., Oman A., Basile D.P., Alvira C.M., Buxton I.L, & Iosef C. (2019). NF‐κB/NKILA signaling modulates the anti‐cancerous effects of EZH2 inhibition. Journal of Cellular and Molecular Medicine, 23(9), 6182-6192.

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Radioligand Binding Assay for RBP4 Interactions

Cited 3 times

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Compound binding to RBP4 was assessed in the radiometric scintillation proximity (SPA) assay that was previously described.^{18 (link),19 (link),26 (link)} The assay measured competitive displacement of radiolabeled [³H]-all-trans-retinol from native RBP4 purified from human urine (Fitzgerald, 30R-AR022L). The protein was biotinylated using the EZ-link Sulfo-NHS-LC-biotinylation kit from ThermoFisher (cat #21335) as recommended by the manufacturer. Binding assays were implemented in a final volume of 100 μL in SPA buffer (1× PBR, pH 7.4, 1 mM ethylenediaminetetraacetic acid (EDTA), 0.1% BRA, 0.5% 3-((3-cholamidopropyl) dimethylammonio)-1-propanesulfonate (CHAPS)). The assay reaction included a radioligand, 10 nM [³H]-all-trans-retinol (48.7 Ci/mmol; PerkinElmer, Waltham, MA), along with the 0.3 mg/well streptavidin-PVT beads (PerkinElmer, RPNQ0006) and 50 nM biotinylated human RBP4. Unlabeled retinol (Sigma-Aldrich, cat #95144) at 20 μM was added to control wells to assess a nonspecific binding. Radioactivity counts were measured using a CHAMELEON plate reader (Hidex Oy, Turku, Finland) after 16 h of incubation at rt with mild shaking.

Cioffi C.L., Muthuraman P., Raja A., Varadi A., Racz B, & Petrukhin K. (2020). Discovery of Bispecific Antagonists of Retinol Binding Protein 4 That Stabilize Transthyretin Tetramers: Scaffolding Hopping, Optimization, and Preclinical Pharmacological Evaluation as a Potential Therapy for Two Common Age-Related Comorbidities. Journal of medicinal chemistry, 63(19), 11054-11084.

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Fluorescence Polarization Assay for XIAP BIR3 Binding

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To determine the binding of a substance to the BIR3-domain of XIAP in vitro we established a FP-assay. Measurements were carried out in black 96-well plates with flat bottom (HVD Life Sciences, Vienna, Austria) in a chameleon plate reader (Hidex, Turku, Finland). 2 μl of substance were added to 103 μl/well XIAP/ARPF Mix containing 20 nM ARPF-K(5-Fam)-NH2-peptide (termed ARPF-FAM) and 720 nM BIR3 protein in assay buffer (100 mM KH2PO4, pH 7.5; 100 μg/ml bovine serum albumin, 0.02% sodium azide). Positive (only assay buffer and FAM-peptide) and negative (BIR3 protein and FAM-peptide) controls were analyzed on each plate. Millipolarization values (mP) were measured at an excitation wavelength of 485 nm and an emission wavelength of 530 nm. Specificity of compound binding was verified using a second FP-assay that measures the interaction between recombinant 14–3–3 sigma protein and R18 peptide [49,50] .

Seiter M.A., Salcher S., Rupp M., Hagenbuchner J., Kiechl-Kohlendorfer U., Mortier J., Wolber G., Rollinger J.M., Obexer P, & Ausserlechner M.J. (2014). Discovery of Sanggenon G as a natural cell-permeable small-molecular weight inhibitor of X-linked inhibitor of apoptosis protein (XIAP). FEBS Open Bio, 4, 659-671.

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