Infinite 1000

Manufactured by Tecan

Sourced in Switzerland, Austria, France

The Infinite 1000 is a compact and versatile microplate reader capable of performing a wide range of absorbance-based assays. It features a xenon flash lamp and filters for wavelengths between 230-1000 nm, allowing users to measure various types of samples, including cells, proteins, and small molecules. The instrument is designed for easy operation and integration into laboratory workflows.

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Lab products found in correlation

Epigenase hdac activity inhibition direct assay kit, by Epigentek (2 mentions) Prestoblue, by Thermo Fisher Scientific (2 mentions) Prestoblue cell viability reagent, by Thermo Fisher Scientific (2 mentions) Vision64, by Bruker (1 mentions) Dektak xt, by Bruker (1 mentions) Axiocam mrc5, by Zeiss (1 mentions) Axiovert 100, by Zeiss (1 mentions) Elisa test, by R&D Systems (1 mentions) Prism 3, by GraphPad (1 mentions) Ficoll 70, by Merck Group (1 mentions) Luciferase reagent, by Promega (1 mentions) Fetal bovine serum (fbs), by Omega Scientific (1 mentions) Griess assay kit, by Beyotime (1 mentions) Apo one homogenous caspase 3 7 kit, by Promega (1 mentions) Prism 8, by GraphPad (1 mentions) Pnpase, by Merck Group (1 mentions) 7 methylguanosine, by Merck Group (1 mentions) Prestoblue cell viability assay, by Thermo Fisher Scientific (1 mentions) Cyquant assay, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Infinite 1000 Pro (7 citations) Infinite 1000 plate reader (3 citations)

24 protocols using infinite 1000

Characterization of PEM Films

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Dry film thickness was measured by using a Dektak XT (Bruker Corporation) profilometer. Three samples (PLL/HA)₂₄ films deposited on silicon were scratched to create a physical step and three measurements per sample were acquired with the software Vision 64® (v 5.4, Bruker Corporation, USA). Scans of 30 s over a length of 1000 μm were performed with a stylus of 12.5 μm in radius and a force set to 1 mg. Thus, dry film thickness is an average value of 9 measurements.
The air dried PEM films coated on PLGA planar substrates were imaged by scanning electron microscopy (SEM) using a FEI-Quanta 250 SEM-FEG in high vacuum at 15 keV using the Everhart-Thornley detector.
The amounts of BMP-2 initially loaded into the PEM films and subsequently released after several washes with the Hepes-NaCl buffer were determined as previously described (1) by fluorescence spectrometry (TECAN Infinite 1000, Austria) using 5% of carboxyfluorescein labeled BMP-2 (BMP-2^CF) as a tracer. Knowing the total surface of the PLGA (see part 2.1), the total surface area of the PEM films was roughly estimated to be 2.6 cm², with the internal and external sides of the PLGA tubes being coated by the film. This value was used to convert the initial and final surface-adsorbed amounts, Γ_I and Γ_R (μg/cm²), respectively, to absolute amounts (μg).

Bouyer M., Guillot R., Lavaud J., Plettinx C., Olivier C., Curry V., Boutonnat J., Coll J.L., Peyrin F., Josserand V., Bettega G, & Picart C. (2016). Surface delivery of tunable doses of BMP-2 from an adaptable polymeric scaffold induces volumetric bone regeneration. Biomaterials, 104, 168-181.

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Evaluating Cell Migration and Invasion

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Cell migration was evaluated by the wound healing assay. Confluent cultures in 6-well plates of NFATc2 shRNA transfectants and control transfectants, or of melanoma cell line 71 were wounded using a sterile 200 μl pipette tip, then washed three times. AM404, 10058-F4, Siomycin A, or GSK126 inhibitors were then added and wound closure was assessed at 48 h by imaging wounds at ×5 through an Axiovert 100 microscope (Zeiss, Oberkochen, Germany) equipped with a digital camera (AxioCam MrC5, Zeiss). Wound width was evaluated through the TScratch software [49 (link)]. Data were expressed as percentage of closure of the original (t = 0 h) wound width. The invasive activity of melanoma cells was tested using the CultrexCoat BME Cell Invasion Assay 96 well (R&D Systems, MN, MN, USA), according to the manufacturer’s instructions. In these experiments melanoma cells were pre-treated for 48 h with different inhibitors as described for the migration assay. Fluorescence signal from calcein-labeled cells was read by an Infinite 1000 instrument (Tecan). Data were expressed as % of invasion by referring to a standard curve.

Perotti V., Baldassari P., Molla A., Nicolini G., Bersani I., Grazia G., Benigni F., Maurichi A., Santinami M., Anichini A, & Mortarini R. (2019). An actionable axis linking NFATc2 to EZH2 controls the EMT-like program of melanoma cells. Oncogene, 38(22), 4384-4396.

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Proliferation Assay of Melanoma Cells

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NFATc2 shRNA and control transfectants were seeded at 8 × 10³ cells/well in 96-well culture plates in the presence of increasing concentrations of FCS. After 144 or 168 h, cultures were evaluated as described [47 (link)] by the 3-(4, 5) dimethylthiazol-2, 5-diphenyltetrazolium bromide (MTT) assay. Effects of Zoledronic acid, GSK126 and their association on melanoma cell proliferation were tested at 72 h. The absorbance was measured at 570 nm with reference at 630 nm, by using an Infinite 1000 instrument (Tecan, Männedorf, Switzerland).

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Quantitative H2S Measurement in Mouse Serum

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The blood samples from each group were collected 12 h after Con A injection, and the blood serums were collected. H₂S concentrations in the serums were evaluated. The mouse serums (75 μL) were mixed with 250 μL of 1% zinc acetate and 425 μL of ddH₂O. Then, 133 μL of 20 mM N-xylylenediamine (7.2 M HCl) and 133 μL of 30 mM FeCl₃ (1.2 M HCl) were added and maintained at room temperature for 10 min. After adding 250 μL of 10% trichloroacetic acid, the samples were centrifuged at 14,000 rpm for 5 min. The supernatant was removed into a 96-well plate, with three duplicates, and the absorbance was measured by a microplate reader (Infinite 1000, TECAN, Switzerland) at OD 670 nm. The calibration curve was linear from 3.125 to 250 µM NaHS.

Ma B., Mao Y., Chang L., Dai T., Xin X., Ma F., Wang Z., Shen Z., Mei Q, & Zhu Y. (2022). S-Propargyl-cysteine prevents concanavalin A-induced immunological liver injury in mice. Pharmaceutical Biology, 60(1), 1169-1176.

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Quantifying SDF-1α Adsorption and Release

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Incorporation and initial release studies were performed on (PLL/HA)₁₂ films constructed in 96-well plates as previously described [34 (link)]. The films were always pre-equilibrated for 30 min in the medium in which SDF-1α was suspended (either 1 mM HCl, or Hepes-NaCl buffer). 50 mL of Alexa-SDF-1α at increasing concentrations from 2.5 to 100 mg/mL were deposited onto the films and adsorbed overnight at 4 °C. 150 mL of Hepes-NaCl were then added in each well and incubated at room temperature for 10 min. The solution was then removed and replaced by fresh Hepes-NaCl buffer before measuring fluorescence (Ex485/Em535) using a fluorescence microplate reader (Infinite1000, Tecan France, Lyon). For release studies, the wells were washed with the Hepes-NaCl solution and the fluorescence was measured at regular time intervals. The incorporated amount was calculated based on a calibration curve obtained with known amounts of Alexa-SDF-1α in solution. SDF-1α loading on film-coated glass slides for cell culture studies was achieved in a similar way, the coated slides being sterilized for 15 min under UV light. Experiments were performed at least in triplicate, with three independent samples per condition in each experiment. Long term SDF-1α release was quantified using an Elisa test (Quantikine, R&D systems, Abingdon, United Kingdom).

Dalonneau F., Liu X.Q., Sadir R., Almodovar J., Mertani H.C., Bruckert F., Albiges-Rizo C., Weidenhaupt M., Lortat-Jacob H, & Picart C. (2014). The effect of delivering the chemokine SDF-1α in a matrix-bound manner on myogenesis. Biomaterials, 35(15), 4525-4535.

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Quantitative Measurement of HDAC Activity

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HDA activity was measured using a commercially available EpigenaseTM HDAC Activity/Inhibition Direct Assay Kit (Epigentek, Cat No. P-4035-48) according to the manufacturer's instruction. 3-17 µl of purified Flag-HDA6 per well were treated with chemicals such as GSNO, GSH, TSA, DTT and incubated with 50 ng of substrate for 90 min at RT. HDA-deacetylated product was immuno-recognized and the fluorescence at 530Ex/590Em nm was measured in a fluorescent microplate reader (Tecan infinite 1000). The RFU values were directly used for relative quantification of HDA activity. HDA activity was also measured according to (Wegener et al., 2003) . 3-17 µl of purified Flag-HDA6 per well were first treated with GSNO or TSA for 30 min in the dark at RT followed by incubation with DTT (if it was required) for another 30 min. The HDA reaction was started by adding 200 µM of HDA-substrate (Boc-Lys(Ac)-MCA) in 25 µl of HDA buffer followed by 60 min incubation at 37°C. The reaction was stopped by adding 45 µl of 2 x Stopping solution containing 10 mg/ml trypsin and 1 µM TSA. The mixture was incubated for an additional 20 min at 30°C to ensure the tryptic digestion. The release of 7-amino-4-methylcoumarin (AMC) was measured by monitoring of florescence at 380Ex/460Em nm.

Ageeva-Kieferle A., Georgii E., Winkler J.B., Ghirardo A., Albert A., Hüther P., Mengel A., Becker C., Schnitzler J., Durner J., & Lindermayr C. (2020). Nitric oxide coordinates histone acetylation and expression of genes involved in growth/development and stress response.

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Spectrophotometric Analysis of Urinary Pyrroles

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Urine samples from the participants were collected in the early morning at the time of physical examination. Samples were stored at −20°C prior to use. Analysis of urinary pyrroles employed minor modifications of published methods.19–21 (link) Pyrrole adducts were measured spectrophotometrically after reaction of 0.08 mL urine with 0.08 mL guanidine hydrochloride (70%) and 0.08 mL of Ehrlich’s reagent 3% 4-dimethylaminobenzaldehyde (DMBA) in the solution of 40% vol/vol methanolic 14% boron trifluoride and 60% vol/vol ethanol.22 (link) Absorption values were measured at 526 nm with an automatic microplate reader (Infinite 1000, Tecan, Switzerland). Calculations were based on a standard curve prepared with different concentrations of 2,5-dimethylpyrrole.

Chen X., Liu W., Wang L., Lin D., Nie L., He K., Guo Z., Zhu F., Feng W., Liu W., Yuan J., Yang X., Spencer P, & Liu J. (2020). Diabetes mellitus is associated with elevated urinary pyrrole markers of γ-diketones known to cause axonal neuropathy. BMJ Open Diabetes Research & Care, 8(1), e001575.

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MTT Assay for Cell Viability Assessment

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Cell viability was assessed by the addition of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolinium bromide (MTT). Briefly, cells were seeded (2–6 × 10⁴ cells/ml) in 96-well plates and allowed to attach for 24 h. Solutions of test compounds were prepared and serially diluted to obtain the appropriate concentrations. The cells were treated with the test compounds at different concentrations (0.032–100 µM) and incubated for 48 h at 37 °C and 5% CO₂. Then, the culture medium was removed, and medium containing 0.2 mg/ml MTT was added. After 3 h (37 °C, 5% CO₂), the MTT-containing medium was removed, and 200 μl of dimethyl sulfoxide (DMSO) was immediately added to each sample. The absorbance was assessed at 540 nm using a Tecan multiplate reader Infinite 1000 (Austria). The half-maximal inhibitory concentration (IC₅₀) of each compound was calculated using Graph Pad Prism 3.0.

Makrecka-Kuka M., Dimitrijevs P., Domracheva I., Jaudzems K., Dambrova M, & Arsenyan P. (2020). Fused isoselenazolium salts suppress breast cancer cell growth by dramatic increase in pyruvate-dependent mitochondrial ROS production. Scientific Reports, 10, 21595.

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MTT Cell Viability Assay Protocol

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Cell viability was based on the ability of cells grown on 96-multiwell plates to convert soluble MTT [3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide] into an insoluble purple formazan reaction product with minor modifications to a previously described protocol [95 (link)]. Briefly, cells were uniformly seeded in 96-multiwell flat bottomed plates and grown in GM. Confluent cultures were washed with PBS and then exposed to DM for 48 h. Afterwards, DM was replaced by RM with or without experimental factors for another 24 h. Relative viability (percentages of mean control value) was calculated. To do this, media were removed and cells were washed with PBS and further incubated with MTT (1 mg/mL) for 1 h at 37 °C in a humidified 5% CO₂ and 95% air in incubator. Next, MTT solution was removed and water insoluble formazan was immediately dissolved in DMSO. The absorbance for MTT was measured at 490 with ELISA reader type Infinite 1000 (TECAN, Mannedorf, Switzerland). Relative percentage (versus untreated controls) of viable cells was measured by MTT conversion into purple formazan.

Pająk B., Kania E., Gołaszewska A, & Orzechowski A. (2019). Preliminary Study on Clusterin Protein (sCLU) Expression in PC-12 Cells Overexpressing Wild-Type and Mutated (Swedish) AβPP genes Affected by Non-Steroid Isoprenoids and Water-Soluble Cholesterol. International Journal of Molecular Sciences, 20(6), 1481.

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Turbidity Assay for Liquid-Liquid Phase Separation

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To analyze LLPS formation using turbidity measurements, Hfq or Hfq-72 in 25 mM tris and 300 mM NaCl (pH 7.5) was diluted 1:10 into 25 mM tris buffer (pH 7.5) supplemented with the indicated concentrations of NaCl, DNA, polyP, and/or 10% (w/v) Ficoll 70 (Sigma-Aldrich) at room temperature (details are given in the figure legends). Turbidity measurements were conducted in 96-well plates at 350 nm using an Infinite1000 (Tecan). All experiments were conducted at least in triplicate.

Beaufay F., Amemiya H.M., Guan J., Basalla J., Meinen B.A., Chen Z., Mitra R., Bardwell J.C., Biteen J.S., Vecchiarelli A.G., Freddolino P.L, & Jakob U. (2021). Polyphosphate drives bacterial heterochromatin formation. Science Advances, 7(52), eabk0233.

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