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96 well plate reader

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The 96-well plate reader is a laboratory instrument designed to measure the optical properties of samples in a 96-well microplate format. It is capable of detecting and quantifying various analytes, such as proteins, nucleic acids, and small molecules, through absorbance, fluorescence, or luminescence detection.

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Lab products found in correlation

Cytotox 96 cytotoxicity kit, by Promega (5 mentions) Celltiter 961 aqueous one solution reagent, by Promega (2 mentions) Tmb substrate, by Thermo Fisher Scientific (1 mentions) Glycerol quantification kit, by Abcam (1 mentions) Cck 8 solution, by Dojindo Laboratories (1 mentions) Celltiter 96 aqueous one solution cell proliferation assay kit, by Promega (1 mentions) Celltiter 96 aqueous one solution cell proliferation assay mts kit, by Promega (1 mentions) Celltiter 96 aqueous one solution cell proliferation assay mts, by Promega (1 mentions) Mtt assay kit, by Cayman Chemical (1 mentions) Softmax pro 5, by Molecular Devices (1 mentions) Mda assay kit, by Beyotime (1 mentions) Prism v8, by GraphPad (1 mentions) Cck 8 kit, by Nanjing Jiancheng (1 mentions) Ab2b10, by Amgen (1 mentions) Ultra tmb substrate, by Thermo Fisher Scientific (1 mentions) Ab2h4 hrp, by Amgen (1 mentions) Huvecs, by American Type Culture Collection (1 mentions) Cell counting kit 8 (cck8), by Solarbio (1 mentions) 96 well plate, by Greiner (1 mentions) Cell titer 96 aqueous solution, by Promega (1 mentions)

40 protocols using 96 well plate reader

1

Colorimetric and Fluorometric Cell Viability Assays

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Cell proliferation was assessed with a standard MTS kit (CellTiter 96 1 AQueous One Solution Reagent (Promega Corp., Madison, WI) according to the manufacturer's recommendations using a 96-well plate reader (Molecular Devices, Sunnyvale, CA). It is a colorimetric method based on the cellular conversion of a tetrazolium compound into a formazan product, which can be detected by the 492 nm absorbance. Such absorbance measurements were not influenced by IR700DX. Cell viability was assessed with a standard alamarBlue Assay (alamarBlue TM Cell Viability Reagent) according to the manufacturer's protocol using a 96-well plate reader (Molecular Devices, Sunnyvale, CA). It is a fluorescent method based on a non-fluorescent molecule resazurin converting to a fluorescent molecule resorufin by reduction reactions in the cytoplasma of metabolically active cells, which can be measured at 590 nm. Such fluorescence measurements were not influenced by IR700DX.
Peng W., de Bruijn H.S., Farrell E., Sioud M., Mashayekhi V., Oliveira S., van Dam G.M., Roodenburg J.L.N., Witjes M.J.H, & Robinson D.J. (2018). Epidermal growth factor receptor (EGFR) density may not be the only determinant for the efficacy of EGFR-targeted photoimmunotherapy in human head and neck cancer cell lines. Lasers in surgery and medicine, 50(5).
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2

Quantifying Cell Cytotoxicity via LDH Assay

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The lactate dehydrogenase (LDH) is a cytoplasmic enzyme retained by viable cells with intact plasma membranes and released from cells with damaged membranes. The LDH release was measured using CytoTox 96 cytotoxicity kit based on the manufacturer’s instructions (Promega, United States) (Zheng et al., 2012 (link)). The levels of maximal LDH release were measured by treating the cultures with a 10× lysis solution (provided by the manufacturer) to yield complete lysis of the cells. Absorbance data were obtained using a 96-well plate reader (Molecular Devices, United States) at 490 nm. According to the manufacturer’s instructions, the LDH release (%) was calculated by calculating the ratio of experimental LDH release to maximal LDH release.
Chen J., Zhuang Y., Zhang Y., Liao H., Liu R., Cheng J., Zhang Z., Sun J., Gao J., Wang X., Chen S., Zhang L., Che F, & Wan Q. (2022). A synthetic BBB-permeable tripeptide GCF confers neuroprotection by increasing glycine in the ischemic brain. Frontiers in Pharmacology, 13, 950376.
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3

ELISA-based Flavivirus Antibody Detection

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Recombinant flavivirus envelope protein was coated on the 96-well plates with the concentration of 1 μg/ml at 4°C overnight. Plates were blocked with 3% BSA in PBS, then incubated with the supernatants or purified mAb in 3-fold serial dilutions. After incubation for 1.5 hours at room temperature, the plates were washed 3 times with PBST (0.5% Tween-20 in PBS), followed by addition of horseradish peroxidase (HRP) coupled goat anti-human IgG (Sigma) and detected using TMB Substrate (Thermo Fisher Scientific) for absorbance signal at OD450 nm using a 96-well plate reader (Molecular Devices). Samples having an OD450 nm of above or equal to 0.1 were designated as positive binders. The OD450 of native control were below 0.4. All assays were replicated three times.
Li L., Meng W., Horton M., DiStefano D.R., Thoryk E.A., Pfaff J.M., Wang Q., Salazar G.T., Barnes T., Doranz B.J., Bett A.J., Casimiro D.R., Vora K.A., An Z, & Zhang N. (2019). Potent neutralizing antibodies elicited by dengue vaccine in rhesus macaque target diverse epitopes. PLoS Pathogens, 15(6), e1007716.
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4

Adrenocortical Inhibition Potency Assay

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The in vivo adrenocortical inhibitory potencies of drugs were assessed in rats using an adrenocorticotropic hormone (ACTH)-stimulation test as previously described. 24 (link) Each rat was given dexamethasone (0.2 mg/kg IV) to suppress baseline corticosterone production. Two hours later, dexamethasone was readministered and the desired dose of drug in dimethyl sulfoxide vehicle was rapidly injected through either a femoral venous catheter preimplanted by the vendor or a 24 gauge intravenous catheter placed in a tail vein. This was followed by a 1-ml normal saline flush. Immediately after the saline flush, ACTH1–24 (25 μg/kg) was injected through the catheter followed by another normal saline flush. Fifteen minutes after ACTH1–24 administration, a blood sample was removed from the catheter (~ 0.4 ml) for measurement of the corticosterone concentration to determine the adrenocortical response to the ACTH1–24. The blood sample was allowed to clot at room temperature before centrifugation at 16000 g for 15 min. The corticosterone concentration in the resulting serum was determined using an Enzyme-Linked ImmunoSorbent Assay (ELISA) (Diagnostic Systems Laboratories, Webster, TX) and a 96-well plate reader (Molecular Devices). For each drug, the median adrenocortical inhibitory dose (ID50) was determined from the dose-corticosterone response relationship using a Hill equation.
Pejo E., Santer P., Jeffrey S., Gallin H., Husain S.S, & Raines D.E. (2014). Analogues of Etomidate: Modifications Around Etomidate’s Chiral Carbon and the Impact on In Vitro and In Vivo Pharmacology. Anesthesiology, 121(2), 290-301.
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5

Oligonol Modulates Adipocyte Lipolysis

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3T3-L1 adipocytes were fully differentiated for 8 days and then treated with different concentration of Oligonol for 24 and 48 hrs. After treatment of differentiated 3T3-L1 cells with Oligonol, free glycerol contents in the cell supernatants were quantified using a glycerol quantification kit according to the manufacturer's instructions (Biovision Inc., Milpitas, USA). Glycerol was quantified at 570 nm on a 96-well plate reader (Molecular devices, Mountain View, USA).
Park J.Y., Kim Y., Im J.A., You S, & Lee H. (2014). Inhibition of Adipogenesis by Oligonol through Akt-mTOR Inhibition in 3T3-L1 Adipocytes. Evidence-based Complementary and Alternative Medicine : eCAM, 2014, 895272.
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6

Keratinocyte Proliferation Assay

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IL-6 KO keratinocytes were pre-treated with IL-6 for 6 hours followed by +/− TGF- β1. Bromodeoxyuridine (BrdU) (EMD, Gibbstown, NJ) was applied immediately following TGF-β1 treatment according to manufacturer's instructions and incubated overnight. Results were obtained via a 96-well plate reader (Molecular Devices, Sunnyvale, CA, U.S.A).
Luckett-Chastain L.R., Cottrell M.L., Kawar B.M., Ihnat M.A, & Gallucci R.M. (2017). Interleukin (IL)-6 modulates transforming growth factor -β receptor I and II (TGF-βRI and II) function in epidermal keratinocytes. Experimental dermatology, 26(8), 697-704.
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7

Cell Viability Assay with CM Treatments

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Cells were plated into 96-well plates (2 × 103 cells/well) and treated with U937/miR-320a-CM, NFs-CM, CAFs-CM, or CAFs/GW4869-CM for 0, 24, 48, and 72 h. After that, 10 µl CCK8 solution (Dojindo, Japan) was added to each pore for 2 hours. Absorbance was recorded at 450 nm with a 96-well plate reader (Molecular Devices Inc., Sunnyvale, CA, USA).
Zhao M., Zhuang A, & Fang Y. (2022). Cancer-Associated Fibroblast-Derived Exosomal miRNA-320a Promotes Macrophage M2 Polarization In Vitro by Regulating PTEN/PI3Kγ Signaling in Pancreatic Cancer. Journal of Oncology, 2022, 9514697.
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8

Cytotoxicity Assay of Sodium Hydrosulfide and Propargylglycine

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The CellTiter 96 AQueous One Solution Cell Proliferation Assay kit (Promega) was used to determine the cytotoxicity of NaHS and PAG in the in vitro studies. Described briefly, 2 × 104 EPCs in 100 μL media were dispensed into each well of 96-well culture plates and cultured overnight. Plates were incubated for 30 min with NaHS (100 μmol/L) or PAG (100 nmol/L). A total of 20 μL of the CellTiter 96 AQueous One Solution Reagent containing tetrazolium compound (3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium, MTS) and an electron coupling reagent (phenazine ethosulfate; PES) was added to each well. Two hours later, the absorbance was recorded at 490 nm using a 96-well plate reader (Molecular Devices, Sunnyvale, CA).
Liu F., Chen D.D., Sun X., Xie H.H., Yuan H., Jia W.P, & Chen A.F. (2014). Hydrogen Sulfide Improves Wound Healing via Restoration of Endothelial Progenitor Cell Functions and Activation of Angiopoietin-1 in Type 2 Diabetes. Diabetes, 63(5), 1763-1778.
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9

Cell Proliferation Assay with MTS

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Cell proliferation was assessed with a standard MTS kit (CellTiter 961 AQueous One Solution Reagent (Promega Corp., Madison, WI, USA) according to the manufacturer’s recommendations using a 96-well plate reader (Molecular Devices, Sunnyvale, CA, USA). The formazan product that is detected by the 492 nm absorbance was not influenced by the IRDye700DX.
Peng W., de Bruijn H.S., ten Hagen T.L., Berg K., Roodenburg J.L., van Dam G.M., Witjes M.J, & Robinson D.J. (2020). In-Vivo Optical Monitoring of the Efficacy of Epidermal Growth Factor Receptor Targeted Photodynamic Therapy: The Effect of Fluence Rate. Cancers, 12(1), 190.
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10

Oligonol Effect on Lipid Release

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After treatment HepG2 cells with a various concentration of Oligonol in the presence of PA, free fatty acid and free glycerol contents in the cell supernatants was quantified using a free fatty acid or glycerol quantification kit according to the manufacturer’s instructions (Biovision Inc., Milpitas, CA, USA). Released free fatty acid or glycerol was quantified at 570 nm on a 96-well plate reader (Molecular devices, Mountain View, CA, USA).
Park J.Y., Kim Y., Im J.A, & Lee H. (2015). Oligonol suppresses lipid accumulation and improves insulin resistance in a palmitate-induced in HepG2 hepatocytes as a cellular steatosis model. BMC Complementary and Alternative Medicine, 15, 185.
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