White opaque 96 well plate

Manufactured by PerkinElmer

Sourced in Singapore

The white opaque 96-well plate is a laboratory equipment used for various assays and experiments. It is a flat, rectangular plate with 96 individual wells, each designed to hold a small volume of liquid sample. The opaque white material of the plate helps to minimize light interference and optimize signal detection during experiments.

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Lab products found in correlation

Envision 2102 multilabel reader, by PerkinElmer (2 mentions) Envision multilabel plate reader, by PerkinElmer (1 mentions) Glosensor 22f plasmid, by Promega (1 mentions) Glosensor camp assay, by Promega (1 mentions) Microbeta2, by PerkinElmer (1 mentions) Optima software, by BMG Labtech (1 mentions) Prism 9, by GraphPad (1 mentions) Celltiter glo, by Promega (1 mentions) Cholesterol cholesterol ester glo assay, by Promega (1 mentions)

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96-well plates (65 citations) White plates (5 citations) Opaque plate (2 citations)

5 protocols using white opaque 96 well plate

Quantifying NF-κB Activity and Inflammatory Markers

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To determine NF-κB activity, in-house developed A549-NF-κB reporter cells [36 (link)] were seeded at a density of 10,000 cells/well in Ham's F-12 medium in white opaque 96-well plates (PerkinElmer). After 24 h, the old medium was replaced with undiluted conditioned medium from nonirradiated or 1- to 3-week-old irradiated fibroblast cultures, and the cells were further incubated for 24 and 48 h. At the end of each incubation, 100 μL Britelite Plus luminescent reagent (PerkinElmer) was added per well, plate content was mixed in a plate shaker, and the luminescent signal was measured in an EnVision Multilabel Plate Reader (PerkinElmer).
The proinflammatory cytokines and/or chemokines in conditioned medium were assayed using a Cytokine Human Magnetic 25-Plex Panel Luminex™ Kit (Life Technologies) following the manufacturer's protocol and analyzed in a Luminex 200 System Analyzer (Austin, TX, USA).
All assays were performed with samples of condition medium obtained from three independent cultures of nonirradiated or irradiated fibroblast cultures.

Agrawal K., Das V., Táborská N., Gurský J., Džubák P, & Hajdúch M. (2018). Differential Regulation of Methylation-Regulating Enzymes by Senescent Stromal Cells Drives Colorectal Cancer Cell Response to DNA-Demethylating Epi-Drugs. Stem Cells International, 2018, 6013728.

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Bioluminescent cAMP Assay for 5-HT1A Receptor

Cited 1 time

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cAMP measurements were performed with a GloSensor cAMP Assay (Promega Corporation, Madison, WI) according to the manufacturer’s protocol. CHO cells were transfected with 7−11 μg of human 5-HT_1AR cDNA and 1 μg of 22F Glosensor plasmid (Promega) using a TransIT-CHO reagent (Mirus Bio, Madison WI), based on established methods.^{39 (link)} Twenty-four hours after transfection, cells were plated in media containing 10% dialyzed FBS and DMEM in white, opaque 96-well plates (PerkinElmer, Waltham, MA) at a density of 150,000 cells/100 μL and incubated at 37 °C, 5% CO₂ overnight. The next day, media were removed from plates, and cells were incubated with 90 μL of assay buffer (1× HBSS +20 mM HEPES) + 1% Glosensor substrate for 2 h in the dark at room temperature. Cells were then treated with compounds made in assay buffer for 30 min in the dark at room temperature. Then, cells were stimulated with 10 μM (final concentration) forskolin for 10 min in the dark at room temperature. Luminescence was measured with a Microbeta^{2 (link)} Microplate Counter (PerkinElmer). Data were analyzed as normalized means (± SEM) from at least three independent determinations, with controls and the kratom alkaloids tested at each concentration in triplicate.

León F., Obeng S., Mottinelli M., Chen Y., King T.I., Berthold E.C., Kamble S.H., Restrepo L.F., Patel A., Gamez-Jimenez L.R., Lopera-Londoño C., Hiranita T., Sharma A., Hampson A.J., Canal C.E., McMahon L.R, & McCurdy C.R. (2021). Activity of Mitragyna speciosa (“Kratom”) Alkaloids at Serotonin Receptors. Journal of medicinal chemistry, 64(18), 13510-13523.

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Caspase-3/7 Activation in Cisplatin-Treated Cells

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The cells were seeded at 20,000 cells/well in a white opaque 96-well plate (PerkinElmer, Singapore) and the plate was left in incubator overnight. Cells were treated with 25 μM cisplatin or vehicle for 24 hrs. Caspase-Gloâ 3/7 Substrate (Promega, USA) was added as per manufacturer's instructions and the plate was incubated at room temperature for 1 hr. Luminescence was measured using a LUMIstar model luminometer (BMG LabTech) using OPTIMA software (BMG LabTech).

Dai Y., Jin S., Li X, & Wang D. (2016). The involvement of Bcl-2 family proteins in AKT-regulated cell survival in cisplatin resistant epithelial ovarian cancer. Oncotarget, 8(1), 1354-1368.

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Cell Viability Assay Optimization

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For cell viability assays, ~1000 mouse colon epithelial cells and ~2500 LIM1215 or SW48 cells were seeded into a white opaque 96-well plate (PerkinElmer). Next day, media was changed to medium supplemented with 2% FBS and inhibitor. Readout of cell proliferation was adopted on cell growth properties avoiding more than 80% confluence in control wells. Cells were then cultured for 72–120 hours depending on the individual doubling times of the cells. Number of living cells were quantified through addition of CellTiter-Glo (Promega) according to the instructions. Plates were read using EnVision 2102 Multilabel Reader (PerkinElmer). EC50 values were calculated in GraphPad Prism 9 based on the point of inflection of a curve.

Budyagan K., Cannon A.C., Chatoff A., Snyder N.W., Kurimchak A.M., Duncan J.S, & Chernoff J. (2024). KRAS mutation-selective requirement for ACSS2 in colorectal adenoma formation. Research Square.

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Intracellular Cholesterol Quantification

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Intracellular cholesterol quantification was performed using Cholesterol/Cholesterol Ester-Glo assay (Promega) according to manufacturer’s instructions. In short, cells were plated into two white opaque 96-well plate (PerkinElmer) and were cultured until ~80% confluency. One of those plates was used for normalization, the other one for cholesterol quantification. For normalization, cell viability was determined using CellTiter-Glo as described above. To measure cholesterol, media was removed and cells were washed twice with PBS. 50 μL of Cholesterol Lysis Solution was added to each well followed by gentle shaking of the plate and incubation for 30 minutes at 37°C. Then, 50 μL of Cholesterol Detection Reagent with or without esterase was added to each well and the plate was shaken for 30–60 seconds at a low rpm on a plate shaker. Cholesterol Detection Reagent without Cholesterol Esterase was used to measure free cholesterol, while with Cholesterol esterase was used to measure total cholesterol. Plates were then incubated for 1 hour at RT. Plates were read using EnVision 2102 Multilabel Reader (PerkinElmer). Cholesterol ester was calculated by subtracting free cholesterol from total cholesterol.

Budyagan K., Cannon A.C., Chatoff A., Snyder N.W., Kurimchak A.M., Duncan J.S, & Chernoff J. (2024). KRAS mutation-selective requirement for ACSS2 in colorectal adenoma formation. Research Square.

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