Ad340 plate reader

Manufactured by Beckman Coulter

Sourced in United States

The Beckman Coulter AD340 plate reader is a versatile instrument designed for absorbance-based detection in microplate applications. The device is capable of measuring absorbance across a wide range of wavelengths, making it suitable for a variety of assays and experiments.

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Lab products found in correlation

Chloramines t, by Merck Group (3 mentions) Ehrlich s solution, by Merck Group (3 mentions) Mtt assay, by Thermo Fisher Scientific (2 mentions) Fast green, by Merck Group (1 mentions) Sirius red, by Merck Group (1 mentions) Chloramine t, by Merck Group (1 mentions) Maxisorp elisa plate, by Thermo Fisher Scientific (1 mentions)

8 protocols using ad340 plate reader

Cytotoxicity Evaluation of Nanoparticles via MTT Assay

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The cytotoxicity of nanoparticles was assessed by MTT assay [3-(4,5-dimethyldiazol-2-yl)2,5 diphenyl Tetrazolium Bromide] [Life Technologies, NY] which quantitatively evaluates the mitochondrial conversion of the MTT salt into purple formazan crystals. Nanoparticle solution was removed, and 0.5 mg/ml MTT working solution in DMEM was incubated on live cells at 37°C for 2.5 h. After incubation, the working solution was removed, and lysis buffer (0.1 N HCl in Isopropanol) added. The lysis buffer was transferred to a 96 well plate and absorbance values collected in an AD340 plate reader [Beckman Coulter, Brea, CA] at corrected 570/620 nm. Relative absorbance was used as the indicator of cell viability. Concentration range of 0 ppm to 1000 ppm for each nanoparticle was used to generate the dose-response curve. SigmaPlot software was used to calculate LC₅₀ value for each type of nanoparticle. Data were expressed as the means ± SD from three independent experiments.

Natarajan V., Wilson C.L., Hayward S.L, & Kidambi S. (2015). Titanium Dioxide Nanoparticles Trigger Loss of Function and Perturbation of Mitochondrial Dynamics in Primary Hepatocytes. PLoS ONE, 10(8), e0134541.

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Cytotoxicity of DOX and HALNP-DOX

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To determine the effect of free DOX and HALNP-DOX on the cell viability of Cort Astro, MG and A172 cells, we utilized an MTT assay as previously reported to calculate the lethal concentration to kill 50% of the cells [57 (link), 74 (link), 75 ]. Following the 24 hour DOX incubation time, the medium of the cells was aspirated, and sterile 5 mg/ml MTT working solution was added and incubated for 2 hours at 37°C. The cells were then lysed with acidified IPA and the absorbance of the produced formazan crystals was measured using a Beckman Coulter AD340 plate reader (Indianapolis, IN, USA). Percent viability was determined by normalization of the 570/620 absorbance ratio to the control untreated cells and positive control dead cells (ethanol treated).

Hayward S.L., Wilson C.L, & Kidambi S. (2016). Hyaluronic acid-conjugated liposome nanoparticles for targeted delivery to CD44 overexpressing glioblastoma cells. Oncotarget, 7(23), 34158-34171.

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Liver Hydroxyproline Quantification

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Liver samples were homogenized in 6 N HCl, hydrolyzed at 100 °C for 18 h, and centrifuged at 9391xg for 5 min. The supernatant was dried in speed-vacuum, dissolved in H₂O, and neutralized with 10 N NaOH. Samples were incubated in a chloramine-T solution [60 mM chloramines-T (Sigma, 857319), 20 mM citrate, 50 mM acetate, pH 6.5] for 25 min at room temperatures, and then in Ehrlich’s solution (Sigma, 038910) at 65 °C for additional 20 min. Hydroxyproline content was measured using a Beckman Coulter AD 340 Plate Reader (570 nm) and normalized to liver weight.

Zhang Z., Zhong X., Shen H., Sheng L., Liangpunsakul S., Lok A.S., Omary M.B., Wang S, & Rui L. (2022). Biliary NIK promotes ductular reaction and liver injury and fibrosis in mice. Nature Communications, 13, 5111.

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Cisplatin Cytotoxicity in OE33 Cells

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OE33 esophageal cancer cells were seeded into 12-well plates at a density of 2 × 10⁴ cells per well for 24 hours before aspiration and subsequent incubation of release media from cisplatin-loaded devices. After 72 hours, in vitro cell viability was measured using the tetrazolium-based colorimetric MTS assay with absorbance at 492 nm with Beckman Coulter AD 340 Plate Reader.

Wang J., Colson Y.L, & Grinstaff M.W. (2017). Tension-activated delivery of small molecules and proteins from superhydrophobic composites. Advanced healthcare materials, 7(7), e1701096.

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Quantifying Hepatic Collagen Content

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Liver paraffin sections were stained with 0.1% Sirius-red (Sigma, 365548) and 0.1% Fast green (Sigma, F7252) (in saturated picric acid). Liver samples were homogenized in 6 N HCl, hydrolyzed at 100°C for 18 h, and centrifuged at 10000 rpm for 5 min. Supernatant was dried in speed-vacuum, dissolved in H₂O, and neutralized with 10 N NaOH. Samples were incubated in a chloramine-T solution (60 mM chloramines-T (Sigma, 857319), 20 mM citrate, 50 mM acetate, pH 6.5) for 25 min at room temperatures, and then in Ehrlich’s solution (Sigma, 038910) at 65°C for additional 20 min. Hydroxyproline content was measured using a Beckman Coulter AD 340 Plate Reader (570 nm) and normalized to liver weight.

Wang P., Kang Q., Wu W.S, & Rui L. (2024). Hepatic Snai1 and Snai2 promote liver regeneration and suppress liver fibrosis in mice. Cell reports, 43(3), 113875.

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Hepatic Hydroxyproline Quantification

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Liver samples were homogenized in 6 N HCl, hydrolyzed at 100 °C for 18 h and centrifuged at 10000 rpm for 5 min. Supernatant was dried in speed-vacuum, dissolved in H₂O, and neutralized with 10 N NaOH. Samples were incubated in a chloramine-T solution (60 mM chloramines-T (Sigma, 857319), 20 mM citrate, 50 mM acetate, pH 6.5) for 25 min at room temperature, and then in Ehrlich’s solution (Sigma, 038910) at 65 °C for additional 20 min. Hydroxyproline content was measured using a Beckman Coulter AD 340 Plate Reader (570 nm) and normalized to liver weight.

Shen H., Sheng L., Xiong Y., Kim Y.H., Jiang L., Chen Z., Liu Y., Pyaram K., Chang C.H, & Rui L. (2017). Thymic NF-κB-inducing Kinase (NIK) Regulates CD4+ T Cell-elicited Liver Injury and Fibrosis in Mice. Journal of hepatology, 67(1), 100-109.

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Quantifying Hepatocyte Viability on Substrates

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The viability of primary hepatocytes on the different substrates was quantified using MTT assay (3-(4,5-dimethylthiazol-2-yl)2,5 diphenyltetrazolium bromide) (Life Technologies, NY). This assay evaluates the mitochondrial conversion of the MTT salt by viable cells to insoluble formazan crystals. Briefly, the cell media was aspirated, and 0.5 mg ml⁻¹ MTT working solution in DMEM was incubated on live cells at 37 °C for 2.5 hours. After incubation, the working solution was removed, and lysis buffer (0.1 N HCl in isopropanol) added to dissolve the purple formazan crystals. Formazan containing lysis buffer was transferred to a 96 well plate and absorbance values collected in an AD340 plate reader [Beckman Coulter, city, state] at corrected 570/620 nm. Relative absorbance was used as the indicator of cell viability.

Natarajan V., Berglund E.J., Chen D.X, & Kidambi S. (2015). Substrate stiffness regulates primary hepatocyte functions. RSC advances, 5(99), 80956-80966.

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Cell Adhesion Assay with Vitronectin

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A 96-well plate (Nunc Maxisorp ELISA plates) was coated with 5 µg/mL vitronectin (50 µL per well) overnight at 4°C. Then, the wells were drained and incubated with 3% (w/v) BSA (50 µL per well) for an hour at 37°C. The negative control wellswere coated with 3% BSA only. The plate was rinsed once with HBSS. 50 µL of 25 000 cells and 50 µL of the LNP per well were added in the wells and incubated at 37°C for 15 min (HEK293(β 3 ) and DU145) or 30 min (PC3). When needed, the cells and LNPs were diluted in HBSS. The plates were drained and gently washed twice with 200 µL of 1X HBSS per well. The surface-bound cells were fixed with ethanol for 15 min at RT and dried at RT or 37°C. The fixed cells were then stained with 1% methylene blue (50 µL per well) for 15 min at RT, rinsed in running waterand dried. The blue dye was finally eluted in 0.1 N HCl (50 µL per well) and the plates were scanned at a 620nm absorbance wavelength using a Beckman Coulter AD340 plate reader. All the measurements were performed in triplicate (or quadruplate) and repeated twice.

Choi J., Rustique E., Henry M., Guidetti M., Josserand V., Sancey L., Boutet J, & Coll J.L. (2017). Targeting tumors with cyclic RGD-conjugated lipid nanoparticles loaded with an IR780 NIR dye: In vitro and in vivo evaluation. International journal of pharmaceutics, 532(2).

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