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96 well half area white plate

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The 96-well half-area white plates are a laboratory equipment product designed for use in various scientific applications. These plates feature a 96-well format with a half-area configuration, providing a reduced sample volume requirement compared to standard full-area plates. The white color of the plates is intended to optimize light reflection and signal detection for applications such as fluorescence and luminescence assays. The plates are made of high-quality materials to ensure durability and consistent performance in the laboratory setting.

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Lab products found in correlation

Victor x3 multilabel plate reader, by PerkinElmer (5 mentions) Firefly luciferase substrate, by Nanolight Technology (2 mentions) Coelenterazine, by Thermo Fisher Scientific (2 mentions) Pfr luc, by Agilent Technologies (1 mentions) Firefly luc assay reagent, by Nanolight Technology (1 mentions) 24 well plate, by Greiner (1 mentions) Glycerol, by Research Products International (1 mentions) Hepes, by Chem-Impex (1 mentions) Ni nta resin, by Thermo Fisher Scientific (1 mentions) Triton x 100, by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Anti sars cov 2 elisa igg, by EUROIMMUN (1 mentions)

7 protocols using 96 well half area white plate

1

Luciferase Assay for Cell Lysis

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After removal of culture medium, cells were frozen at −80 °C to ensure efficient cell disruption. Cells were lysed in 100 µl of lysis buffer (20 mM Tris-HCl, 10% glycerol, 0.1% Triton X-100, 1 mM PMSF, 0.1% β-mercaptoethanol, pH 8.0) for 30 min at room temperature on swinging platform shaker. Luciferase assay was performed in 96-well half-area white plates (Costar) by mixing 10 μl of cell lysate with 20 μl of firefly luciferase substrate (Nanolight Technology). Bioluminescence was measured immediately with a Victor X3 multilabel plate reader (Perkin Elmer), and data were analyzed with an OriginPro 8.6 software.
Leopold A.V., Chernov K.G., Shemetov A.A, & Verkhusha V.V. (2019). Neurotrophin receptor tyrosine kinases regulated with near-infrared light. Nature Communications, 10, 1129.
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2

Luciferase Assay of Light-Induced Cells

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PC6–3 and HeLa cells transfected and kept under 660 nm (0.5 mW cm−2) or 780 nm (0.5 mW cm−2) light as described above were then lysed in 100 μl of lysis buffer (20 mM Tris-HCl, 10% glycerol, 0.1% Triton X-100, 1 mM PMSF, 0.1% β-mercaptoethanol, pH 8.0) for 30 min at room temperature on swinging platform shaker. Luciferase assay was performed in 96-well half-area white plates (Costar) by mixing 10 μl of cell lysate with 20 μl of firefly luciferase substrate (Nanolight Technology). Bioluminescence was measured immediately with a Victor X3 multilabel plate reader (Perkin Elmer), and data were analyzed with an OriginPro v. 8.6 software.
Leopold A.V., Pletnev S, & Verkhusha V.V. (2020). Bacterial phytochrome as a scaffold for engineering of receptor tyrosine kinases controlled with near-infrared light. Journal of molecular biology, 432(13), 3749-3760.
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3

Light-Controlled Gene Expression Activation

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For light-controlled transcription activation, cells were cotransfected with the constructs pQP-T2A[11 (link)] (Addgene #102583) and pFR-Luc (Agilent Technologies) in a 3:1 ratio in 24-well plates (Greiner Bio-One). Illumination of plates was applied directly in CO2 incubator with 740/25 nm light (0.2 mW cm−2) in alternating cycles: 30 s light and 180 s darkness. To measure Fluc activity, transfected cells were lysed 48 h after transfection. Cells were washed with PBS, and then 100 μl of lysis buffer (20 mM Tris–HCl pH 8.0, 10% glycerol, 0.1% β-mercaptoethanol, 0.1% Triton X-100, 1 mM PMSF) was added to each well of 24-well plate and incubated on ice for 30 min. 10 μl of cell lysates were mixed with 20 μl of Firefly Luc Assay reagent (NanoLight Technology) in 96-well half-area white plates (Costar). Bioluminescence signal was measured using a Victor X3 multilabel plate reader (PerkinElmer).
Redchuk T.A., Karasev M.M., Omelina E.S, & Verkhusha V.V. (2018). Near-infrared light-controlled gene expression and protein targeting in neurons and non-neuronal cells. Chembiochem : a European journal of chemical biology, 19(12), 1334-1340.
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4

Optimized UMP-Glo Assay Protocol

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The prototype UMP-Glo assay kit was a generous gift from Promega. HEPES (99.9%) was obtained from Chem-Impex International. NaCl (>99%) and glycerol (>99%) were from Research Products International. Triton X-100 and DMSO (99.9%) were from Sigma-Aldrich, MgCl2.6H2O (100%) was from Mallinckrodt Chemicals. DDM (>99%) was purchased from Anatrace. 96-well half-area white plates were obtained from Corning. Ni-NTA resin was from Thermo Fisher. All other chemicals and bio-reagents were purchased at the purest grade available.
Das D., Walvoort M.T., Lukose V, & Imperiali B. (2016). A Rapid and Efficient Luminescence-based Method for Assaying Phosphoglycosyltransferase Enzymes. Scientific Reports, 6, 33412.
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5

SARS-CoV-2 Spike Pseudovirus Neutralization Assay

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Convalescent serum from COVID-19 patients or healthy donors (collected before 30 April 2020) was obtained from Children’s Hospital Los Angeles. Convalescent sera were confirmed to be positive for IgG class antibodies against SARS-CoV-2 Spike using anti-SARS-CoV-2 ELISA (IgG) (Euroimmun, Lübeck, Germany) (78 ).
A suitable dose of Spike-pseudotyped VSV-Luc vectors was used in the neutralization assays to produce approximately 105 relative light units (RLU) of luciferase activity on HeLa-ACE2 cells in the absence of serum. A total of 5 × 103 HeLa-ACE2 cells were seeded in tissue culture-treated, 96-well half-area white plates (Corning) to achieve 50% to 75% confluence the following day. Convalescent or control sera were 3-fold serially diluted from 1:10 to 1:7,290, and 50 μl was incubated with the predetermined dose of the VSV-Luc vectors for 30 min at 37°C before addition of the mixture to HeLa-ACE2 cells. Cells were incubated at 37°C overnight for 16 to 24 h. Vector transduction efficiency was quantified by measuring luciferase activity as described above and neutralization (%) calculated by normalization to the values obtained on cells transduced without serum.
Chen H.Y., Huang C., Tian L., Huang X., Zhang C., Llewellyn G.N., Rogers G.L., Andresen K., O’Gorman M.R., Chen Y.W, & Cannon P.M. (2021). Cytoplasmic Tail Truncation of SARS-CoV-2 Spike Protein Enhances Titer of Pseudotyped Vectors but Masks the Effect of the D614G Mutation. Journal of Virology, 95(22), e00966-21.
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6

Antigen-Dependent Transcriptional Activation Assay

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For an antigen-dependent GAL4 stabilization, HeLa cells were co-transfected with plasmids encoding NIR-FbGFP–GAL4 or biNIR-FbGFP–GAL4, reporter plasmid pUAS-Gluc (5x UAS), and with either EGFP or mTagBFP in a 1:10:89 ratio. HeLa cells were co-transfected with plasmids encoding ALFA-tagged GAL4-VP16, reporter plasmid pUAS-Gluc (5x UAS), NIR-FbGFP–NbALFA or biNIR-FbGFP–NbALFA, and with either EGFP or mTagBFP2 in the 1:10:44.5:44.5 ratio. As controls, cells transfected with reporter plasmid pUAS-Gluc and with or without ALFA-tagged GAL4-VP16 were used. To measure Gluc activity, the co-transfected HeLa cells were grown in 24-well plates. 10 h after the transfection mixture was removed, a fresh growth medium was added, and cells were incubated for 48 h. Then, 5 μl of culture media was mixed with 100 μl of 2 μM coelenterazine (Invitrogen/Thermo Fisher Scientific) in PBS in wells of a 96-well half-area white plate (Costar). Bioluminescence was measured using a Victor X3 multilabel plate reader (PerkinElmer).
Oliinyk O.S., Baloban M., Clark C.L., Carey E., Pletnev S., Nimmerjahn A, & Verkhusha V.V. (2022). Single-domain near-infrared protein provides a scaffold for antigen-dependent fluorescent nanobodies. Nature methods, 19(6), 740-750.
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7

Antigen-Dependent Transcriptional Activation Assay

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For an antigen-dependent GAL4 stabilization, HeLa cells were co-transfected with plasmids encoding NIR-FbGFP–GAL4 or biNIR-FbGFP–GAL4, reporter plasmid pUAS-Gluc (5x UAS), and with either EGFP or mTagBFP in a 1:10:89 ratio. HeLa cells were co-transfected with plasmids encoding ALFA-tagged GAL4-VP16, reporter plasmid pUAS-Gluc (5x UAS), NIR-FbGFP–NbALFA or biNIR-FbGFP–NbALFA, and with either EGFP or mTagBFP2 in the 1:10:44.5:44.5 ratio. As controls, cells transfected with reporter plasmid pUAS-Gluc and with or without ALFA-tagged GAL4-VP16 were used. To measure Gluc activity, the co-transfected HeLa cells were grown in 24-well plates. 10 h after the transfection mixture was removed, a fresh growth medium was added, and cells were incubated for 48 h. Then, 5 μl of culture media was mixed with 100 μl of 2 μM coelenterazine (Invitrogen/Thermo Fisher Scientific) in PBS in wells of a 96-well half-area white plate (Costar). Bioluminescence was measured using a Victor X3 multilabel plate reader (PerkinElmer).
Oliinyk O.S., Baloban M., Clark C.L., Carey E., Pletnev S., Nimmerjahn A, & Verkhusha V.V. (2022). Single-domain near-infrared protein provides a scaffold for antigen-dependent fluorescent nanobodies. Nature methods, 19(6), 740-750.
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