Cells were lysed in 1 × passive lysis buffer, scraped, pelleted and then assayed for luciferase and Renilla activity (Promega) according to the manufacturer's instructions and using the Synergy 2 plate reader (Bioteck, Winooski, VT).
Synergy 2 plate reader
Manufactured by Bioteck
Sourced in Japan
The Synergy 2 plate reader is a multi-mode microplate reader designed for a variety of common laboratory assays. It utilizes advanced optical components and monochromator-based technology to provide accurate and reliable detection across multiple detection modes, including absorbance, fluorescence, and luminescence.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using synergy 2 plate reader
1
Luciferase and Renilla Assay Protocol
2
Quantifying Phospho-AKT and mTORC1 Activity
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Endogenous levels of phosphorylated AKT at Ser473 were determined with FastScan™ Phospho‐Akt (Ser473) ELISA Kit (Cell Signaling technologies, CST#80895) in cleared cell lysates of mock treated and virus infected 106 A549 cells. Absolute values of phosphorylated Ser473‐AKT were determined with a standard curve correlating the positive control phospho‐Akt(Ser473) concentration (μg/ml) to absorbance at 450 nM. We utilized PathScan Phospho‐p70 S6 Kinase (Thr389) Sandwich ELISA kit (Cell Signaling technologies, CST#7063) to determine mTORC1 activity by measuring the phosphorylated levels of its primary substrate p70S6K. In this assay, the levels of phosphorylated Thr389 (detection antibody) were normalized to the total levels of p70S6K protein (capture antibody). A Bioteck Synergy 2 plate reader was used to collect absorbance at 450 nm.
3
Measuring Aβ₄₂ Peptide Solubility
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The preformed Cu2+-free and -bound Aβ42 aggregates were treated with FC-11 or FC-11-1 in darkness, or exposed to UV light, or treated by both for 0–4 h in pH 7.4 buffer at 37 °C. Following this treatment, the supernatants were collected by centrifugation for 20 min at 14,000 rpm. The absorbance was measured at 562 nm for all the supernatants with a Bioteck synergy-2 plate reader. The freshly prepared Aβ42 solution was used as control. The soluble Aβ42 peptide content in the supernatants was assayed by a BCA protein assay kit using bovine serum albumin as standard.
4
Fluorescence Polarization Assay for LIG1-TTD Binding
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Fluorescence polarization assays for interaction between LIG1 peptide and TTD were performed in binding buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 10% glycerol, 1 mM DTT, and 0.05% Tween 20) at 25 C using a Synergy2 plate reader (Bioteck Japan). The excitation and emission wavelengths were 485 nm and 522 nm, respectively. The 6-Carboxyfluorescein Hydrate (FAM) labeled LIG1 peptide (10 nM) was incubated with increasing concentrations of the TTD. Curve-fitting analyses and dissociation constant (K d ) were conducted using ORIGIN software version 8.0 (OriginLab). The observed data were fitted to the equations assuming a 1:1 binding stoichiometry ratio.
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