Cells were first treated with 10 μM of LVV-H7 for 30 min at 37 °C. Then, the Nanoluc substrate furimazine (10 μM) was added and BRET signals were immediately measured at 37 °C before and after adding 100 nM of BODIPY-AngII using the CLARIOstar multilabel plate reader (BMG Labtech (Australia)) as described above.
Clariostar multilabel plate reader
Manufactured by BMG Labtech
Sourced in Australia
The CLARIOstar is a multilabel plate reader designed for sensitive, high-performance detection of a wide range of assays. It features state-of-the-art optics and advanced detection technologies to provide reliable and accurate results.
Automatically generated - may contain errors
3 protocols using clariostar multilabel plate reader
1
BRET Assay for AngII Binding
2
Evaluating AVNP Cytotoxicity in MARC-145 Cells
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AVNP suspensions were serially diluted in cDMEM and added to MARC-145 cells in 96 well white-walled plates (Fisher Scientific, Loughborough, UK). After 24 h incubation, cytotoxicity was assessed using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega, Southampton, UK). Following the addition of substrate, luminescence was measured using a CLARIOstar multilabel plate reader (BMG LabTech, Aylesbury, UK).
3
BODIPY-AngII Binding Assay
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Cells were first treated with olmesartan medoxomil, LVV-H7 and/or vehicle (as described in figure legends) for 20 min at 37 °C followed by addition of increasing doses of BODIPY-AngII and incubation for a further 40 min at 37 °C. The NanoLuc substrate furimazine (Promega) was then added to a final concentration of 10 μM and light emission was measured at 37 °C using the filters 450 nm (80-nm bandpass) and >610 nm (longpass) on the LUMIstar or CLARIOstar multilabel plate reader (BMG Labtech (Australia), Mornington, VIC, Australia).
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