Victor x3 multilabel plate reader

Manufactured by PerkinElmer

Sourced in United States, Italy

The VICTOR X3 Multilabel Plate Reader is a versatile microplate reader that can perform a variety of detection methods, including absorbance, fluorescence, and luminescence. It is designed to provide accurate and reliable data for a wide range of applications in life science research and drug discovery.

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Victor X3 (440 citations) Victor plate reader (111 citations) Multilabel plate reader (45 citations) Victor X3 reader (16 citations)

179 protocols using victor x3 multilabel plate reader

Embryonic DNA Methylation Assay

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Newly fertilized
eggs were collected
immediately after spawning and placed in groups of approximately 100
per Petri dish within a light- and temperature-controlled incubator.
Embryos (100 per time point) were collected at 2, 10, and 24 hpf and
stored at −20 °C. Nuclear proteins were extracted from
whole embryo pools using an EpiQuik Nuclear Extraction Kit (Epigentek
Group, Farmingdale, NY). Nuclear extract was kept on ice and immediately
quantified using a BCA Protein Assay (Pierce Biotechnology, Rockford,
IL) following the manufacturer’s instructions. Optical density
of the colorimetric reaction was quantified using a VICTOR X3Multilabel
Plate Reader (PerkinElmer, Waltham, MA), and total protein was quantified
using a standard curve generated from bovine serum albumin.
DNMT inhibition was quantified using an EpiQuik DNMT Activity/Inhibition
Assay Ultra Fluorometric Kit (Epigentek Group, Farmingdale, NY). DNMT
inhibition within nuclear extracts (6.5 μg of protein per reaction)
derived from 2-, 10-, and 24-hpf embryos was quantified in the presence
of vehicle (0.1% dimethyl sulfoxide, DMSO), 250 μM 5-azaC, or
TDCIPP (63, 125, 250, or 500 μM). All reactions were conducted
in triplicate. Fluorescence was measured by a VICTOR X3Multilabel
Plate Reader (PerkinElmer, Waltham, MA) and data were corrected for
background and reported as relative fluorescent units.

Volz D.C., Leet J.K., Chen A., Stapleton H.M., Katiyar N., Kaundal R., Yu Y, & Wang Y. (2016). Tris(1,3-dichloro-2-propyl)phosphate Induces Genome-Wide Hypomethylation within Early Zebrafish Embryos. Environmental Science & Technology, 50(18), 10255-10263.

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WST-1 and PicoGreen Assay for Cell Proliferation

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For
WST-1 assay, cells
were seeded in a 24-well cell culture plates (COSTAR) at a density
of 1.5 × 10⁴ cells/well. After 24 h, cells were incubated
with IDE-NLC preparations at different IDE molar concentrations in
complete DMEM without phenol red (Sigma-Aldrich). In parallel, cells
were incubated with free IDE dissolved in ethanol and NLCs. Studies
were performed at 24 and 72 h after incubation at 37 °C, 5% CO₂. Dilution (1:20) of cell proliferation reagent WST-1 (Roche)
was prepared in 300 μL/well of DMEM without phenol red, and
plates were incubated at 37 °C for 30 min. Absorbance reading
was performed with a Victor X3 Multilabel Plate Reader (Perkin Elmer),
setting absorbance wavelength at 450 nm and 0.1 s measurement time.
Cells were then rinsed with PBS and frozen at −80 °C.
Frozen samples were treated to assess the proliferation rate by Quant-iT
PicoGreen dsDNA Assay Kit (Invitrogen). Samples stocked at −80
°C were subjected to three cycles of freeze/thaw in order to
allow cell lysis and DNA release. Meanwhile, a series of dilutions
of the reagent and buffer were prepared according to the manufacturer’s
instructions. Fluorescence was measured with Victor X3 Multilabel
Plate Reader (Perkin Elmer), setting the excitation wavelength at
485 nm, emission wavelength at 535 nm, and 0.1 s measurement time.

Martinelli C., Battaglini M., Pucci C., Gioi S., Caracci C., Macaluso G., Doccini S., Santorelli F.M, & Ciofani G. (2020). Development of Nanostructured Lipid Carriers for the Delivery of Idebenone in Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay. ACS Omega, 5(21), 12451-12466.

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Antioxidant Capacity Evaluation of Fruit Extracts

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The DPPH• radical scavenging activity of FS and NFS extracts was determined according to Chiellini et al. [40 (link)]. The absorbance was recorded at 517 nm, and the extract concentration corresponding to 50% of DPPH inhibition (EC₅₀) was measured according to Gabriele et al. [41 (link)]. The oxygen radical absorbance capacity (ORAC) of FS and NFS extracts was determined as described by Gabriele et al. [39 ]. AAPH was used as a peroxyl radicals generator and fluorescein as the probe. Fluorescein fluorescence decay was read at λ_ex 485 nm and λ_em 514 nm using a VictorTM X3 Multilabel Plate Reader (Perkin Elmer, Waltham, MA, USA). Results were expressed as ORAC units (µmol TE/g FW) using Trolox as the reference standard. The ferric-reducing antioxidant power (FRAP) assay was used to evaluate the ability of FS and NFS extracts to reduce ferric iron (Fe³⁺) to ferrous iron (Fe²⁺) [42 (link)]. The absorbance was measured at 593 nm (Perkin Elmer UV/VIS Lambda 365, Waltham, MA, USA), and results were expressed as Fe²⁺ equivalents (µM) using a standard curve of FeSO₄·7H₂O. The Fe²⁺ chelation ability of FS and NFS extracts was determined as described by Chelucci et al. [42 (link)]. The absorbance was read at 562 nm (Perkin Elmer UV/VIS Lambda 365, Waltham, MA, USA), and results were expressed as EC₅₀ values referring to the extract concentration corresponding to 50% of Fe²⁺ chelation.

Gabriele M., Arouna N., Árvay J., Longo V, & Pucci L. (2023). Sourdough Fermentation Improves the Antioxidant, Antihypertensive, and Anti-Inflammatory Properties of Triticum dicoccum. International Journal of Molecular Sciences, 24(7), 6283.

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Quantifying Cytoprotective Effects of H2S

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HL-1 cells were seeded in 12-well plates at 10,000 cells per well in complete Claycomb medium. Cells were preconditioned with GYY4137 over 12 hours and ATP content was analyzed periodically. To evaluate the cardio-protective effect of H₂S during nutrient deprivation, HL-1 cells were preconditioned overnight with GYY4137 and the culture medium was then replaced with PBS to promote starvation. ATP content was analyzed at various times. To measure ATP, cells were washed twice with PBS buffer and then lysed with the addition of somatic cell ATP releasing reagent (FLSAR, Sigma-Aldrich). The supernatant was collected and kept at −20°C until analysis. ATP content was measured with the bioluminescent ATP somatic cells assay kit (FLASC, Sigma-Aldrich). Luminescence of triplicate samples was read on a VICTORTM X3 Multilabel Plate Reader (PerkinElmer) with a test wavelength of 570 nm. Rat hearts were explanted and stored at −80°C for ATP quantification with the colorimetric ATP Assay Kit (Abcam) following the manufacturer’s instructions. Briefly, tissue samples were homogenized in ATP assay buffer, deproteinized with 1 M perchloric acid, and ATP content was determined by absorbance at 570 nm. ATP levels from experimental samples were compared to sham samples (explanted hearts without treatment).

Garcia N.A., Moncayo-Arlandi J., Vazquez A., Genovés P., Calvo C.J., Millet J., Martí N., Aguado C., Knecht E., Valiente-Alandi I., Montero J.A., Díez-Juan A, & Sepúlveda P. (2017). Hydrogen Sulfide Improves Cardiomyocyte Function in a Cardiac Arrest Model. Annals of Transplantation, 22, 285-295.

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Assessing Migration, Invasion, and Proliferation of Glioma Cells

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After stable overexpression LN229 and U343 and knockdown U87 cells reached ~70% confluence, the cells were scratched with a 200 µL pipette tip, washed with DPBS, and incubated at 37 °C. Wound healing was observed for 12 h and 24 h at the scratched line, and the area around the scratched line was photographed. The invasion assay was performed using an 8.0 μm pore polycarbonate membrane insert (Corning, 353097). Transwell inserts coated with Matrigel (Corning, 354234) were used, and 2 × 10⁴ cells/well were plated in the upper chamber. The lower chamber was filled with 600 μL of serum-free medium. After incubation for 48 h, cells infiltrated with 100% methanol were fixed. After staining with 1% crystal violet (Sigma‒Aldrich, V5265), the infiltrated cells were observed under a microscope. For the colony formation assay, 1 × 10³ cells were plated in 6-well plates. After culturing the cells at 37 °C for 2 weeks, colonies formed on each of the three plates were measured. Cell proliferation was assessed according to the manufacturer’s protocol (Invitrogen, C35011) using the CyQuant Direct Cell Proliferation Assay Kit. All experiments were performed in triplicate, and cell proliferation was determined using a VICTOR^TM X3 Multilabel Plate Reader (PerkinElmer).

Jeong J.H., Park S.H., Kim H., Nam H.Y., Kim S.H., Jeong M., Kong M.J., Son J., Jeong J.E., Song J.H., Kim S.W, & Choi K.C. (2023). ZBTB7A suppresses glioblastoma tumorigenesis through the transcriptional repression of EPB41L5. Experimental & Molecular Medicine, 55(1), 43-54.

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Cell Viability Assay with Sorafenib and Etoposide

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For the cell viability assays, the cells were seeded in 96-well plates at a density of 5000 cells/well (100 μL total volume/well) and were grown for 24 h. The cells were then washed with sterile phosphate-buffered saline and maintained in low-glucose (5.56 mM) DMEM supplemented with 10% FBS with/without sodium pyruvate (1 mM, Thermo Fisher Scientific, Waltham, MA, USA) for 24 h before adding the sorafenib (LC Laboratories, Woburn, MA, USA) or etoposide (Sigma-Aldrich, Saint Louis, MO, USA). The cells were treated with different concentrations of sorafenib or etoposide, and the controls were treated with vehicle (DMSO, Sigma-Aldrich). After 24-h treatment, WST-1 solution (DoGenBio, Seoul, South Korea) was added to the cells for 1–2 h, and the absorbance at 450 nm was then measured using a VICTORTMX3 Multilabel Plate Reader (PerkinElmer Inc., Waltham, MA, USA).

Kim D., Hwang C.Y, & Cho K.H. (2024). The fitness trade-off between growth and stress resistance determines the phenotypic landscape. BMC Biology, 22, 62.

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Cytocompatibility Evaluation of NETA-Loaded Biomaterials

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Cytocompatibility of the drug-free and NETA-loaded segments was evaluated (n = 5) using an MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide dye). For this assay, a cell suspension with density of 2 × 10⁴ cells/well in 100 µL was seeded on 96 well plates and incubated for 24 h at 37 °C and 95% relative humidity containing 5% CO₂. Afterward, the growth medium was aspirated and replaced with a growth medium consisting of the extracted leachates, with the cells thereafter grown for an additional 1, 3, 5 and 7 days. Throughout the 7-day testing period, the growth medium containing the extracted leachates was replenished every 2 days. At the end of each designated time point (1, 3, 5, and 7 days), MTT solution (10 µL) was added to each well plate, followed by an addition of 100 µL solubilizing buffer after 4 h, and incubation overnight under the standard culture conditions. The absorbance’s reading of each treatment sample was thereafter taken in triplicate at 570 nm against a 690 nm background using a VICTORTM X3 Multilabel Plate Reader (PerkinElmer, Waltham, MA, USA). In addition to the treatment arms (the NETA-loaded and drug-free segments), NIH/3T3 cells in culture medium and cells in culture medium with DMSO were used as negative and positive controls, respectively.

Abdelgader A., Govender M., Kumar P, & Choonara Y.E. (2024). A Novel Intrauterine Device for the Spatio-Temporal Release of Norethindrone Acetate as a Counter-Estrogenic Intervention in the Genitourinary Syndrome of Menopause. Pharmaceutics, 16(5), 587.

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Quantifying Cellular Reactive Oxygen Species

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ROS were quantified using 2′,7′-dichlorofluorescin-diacetate (H₂-DCF-DA, Sigma-Aldrich, St. Louis, MO, USA), as previously described [17 (link)]. Upon cleavage of the acetate groups by intracellular esterase and oxidation, the H₂−DCF-DA is converted to the fluorescent 2′,7′-dichlorofluorescein (DCF). Briefly, the intestinal cells were seeded into 96-well plates and allowed to adhere overnight. ROS level was measured after the exposure to BOS (1 µg/mL) or to CUR (1 µg/mL) for 24 h. Treatments were removed and H₂DCF-DA was added to obtain a final concentration of 50 µM in each well. The plate was incubated for 30 min at 37 °C and washed with phosphate-buffered saline (PBS). DCF fluorescence intensity was measured at excitation 485 nm–emission 535 nm, using VICTOR^TMX3 Multilabel Plate Reader (PerkinElmer, Waltham, MA, USA), before and after the addition of 500 µM H₂O₂ on each well.

Governa P., Marchi M., Cocetta V., De Leo B., Saunders P.T., Catanzaro D., Miraldi E., Montopoli M, & Biagi M. (2018). Effects of Boswellia Serrata Roxb. and Curcuma longa L. in an In Vitro Intestinal Inflammation Model Using Immune Cells and Caco-2. Pharmaceuticals, 11(4), 126.

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Assessing Adenoviral Transduction in Murine Cells

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Human cells were seeded in 96-well plates (1 × 10⁴ cells/well) and incubated overnight at 37 °C and 5% CO₂. Fresh serum from naïve C57BL/6 mice, Rag2^-/-, or Ighm^tm1Che C57BL/6 mice was incubated with 1 × 10⁹ vp of vector (HAdV-5 luc) in a final volume of 50 μL (30 min at 37 °C). The potential interactions with mouse FX were inhibited by the addition of 40 μg/mL X-bp. Samples were then diluted 200-fold in serum-free medium. A total of 1000 vp/cell were added onto cells for 2 h at 37 °C, then media was replaced with fresh media containing 2% FBS, and after further ~16 h at 37 °C cells were rinsed with PBS and harvested by freezing and thawing in Reporter Lysis Buffer (Promega, UK) for both luciferase assay (Promega, Southampton, UK) and protein content measurement (BCA assay) using a VICTOR^TM X3 Multilabel Plate Reader (PerkinElmer, Waltham, MA, USA).

Doszpoly A., de la Cuesta F., Lopez-Gordo E., Bénézech C., Nicklin S.A, & Baker A.H. (2019). Human Adenovirus Serotype 5 Is Sensitive to IgM-Independent Neutralization In Vitro and In Vivo. Viruses, 11(7), 616.

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Labeling and Harvesting Cell Sheets

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Prior to cell sheet formation MSC (passage 2–4) were labeled by PKH26 Red dye (PKH26 Red Fluorescent Cell Linker Kit for General Cell Membrane Labeling, Sigma-Aldrich, Milwaukee, WI, USA). The cells were detached with 0.05% trypsin, labeled and seeded at 3 × 106/well density in a 6-well culture plate (Corning, Corning, NY, USA). Forming CSs were maintained in DMEM, 10% FBS, penicillin/streptomycin for at least 48 h prior to experiments. The transgene expression was verified by enzyme-linked immunosorbent assay to assess SCF concentration in conditioned media. The Mouse SCF ELISA Kit (ab100740, Abcam, Cambridge, MA, USA) was used. An optical density was measured using VictorTM X3 Multi Label Plate Reader (Perkin Elmer Inc, Waltham, MA, USA). The cell sheet was harvested by incubation in Versene solution (Paneco, Moscow, Russia) until it self-detached within 5–7 min.

Dergilev K.V., Shevchenko E.K., Tsokolaeva Z.I., Beloglazova I.B., Zubkova E.S., Boldyreva M.A., Menshikov M.Y., Ratner E.I., Penkov D, & Parfyonova Y.V. (2020). Cell Sheet Comprised of Mesenchymal Stromal Cells Overexpressing Stem Cell Factor Promotes Epicardium Activation and Heart Function Improvement in a Rat Model of Myocardium Infarction. International Journal of Molecular Sciences, 21(24), 9603.

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