NanoLuc® Binary Technology (NanoBiT) assays were performed with 2×105 cells/ml HEK293T cells transiently transfected with 1 μg/ml of the N-LgBiT-RING1B plasmid (full length RING1B or RING1B R98A cloned into pBiT1.1-N[TK LgBiT vector, Promega) and 0.25 μg/ml of the SmBiT-H3.3 plasmid (CMV/H3.3-SmBiT vector, Promega), and 7:1 FuGENE® HD Transfection Reagent-to-DNA ratio (Promega) in Opti-MEM™ (Gibco). After 24 h, cells were trypsinized and transferred to white 96-well tissue culture plates (30,000 cells in 100 μl per well) and incubated with compounds or 0.25% DMSO. After another 24 h, 25 μl of Nano-Glo® Live Cell Assay System (Promega) were added to each well and NanoLuc® Luminescence was detected in a PHERAstar BMG microplate reader.
Cmv h3.3 smbit vector
Manufactured by Promega
The CMV/H3.3-SmBiT vector is a genetic engineering tool used for the expression and detection of target proteins. It contains a cytomegalovirus (CMV) promoter for driving gene expression and the H3.3 histone variant fused to the small BioBit (SmBiT) tag. This vector enables the monitoring of target protein localization and interactions in live cells.
Automatically generated - may contain errors
2 protocols using cmv h3.3 smbit vector
1
NanoLuc Binary Technology (NanoBiT) Assay
2
NanoLuc Binary Technology (NanoBiT) Assay
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NanoLuc® Binary Technology (NanoBiT) assays were performed with 2×105 cells/ml HEK293T cells transiently transfected with 1 μg/ml of the N-LgBiT-RING1B plasmid (full length RING1B or RING1B R98A cloned into pBiT1.1-N[TK LgBiT vector, Promega) and 0.25 μg/ml of the SmBiT-H3.3 plasmid (CMV/H3.3-SmBiT vector, Promega), and 7:1 FuGENE® HD Transfection Reagent-to-DNA ratio (Promega) in Opti-MEM™ (Gibco). After 24 h, cells were trypsinized and transferred to white 96-well tissue culture plates (30,000 cells in 100 μl per well) and incubated with compounds or 0.25% DMSO. After another 24 h, 25 μl of Nano-Glo® Live Cell Assay System (Promega) were added to each well and NanoLuc® Luminescence was detected in a PHERAstar BMG microplate reader.
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