A N-terminal NanoLuc® luciferase ID1 fusion protein was synthesized by Genewiz (South Plainfield, NJ, USA) into the pUC57 backbone and then cloned into the pcDNA3.1 plasmid with EcoRI-HF and XbaI (both enzymes from New England Biolabs, Ipswich, MA, USA). The assay was performed essentially as described in the NanoBRET™ TE Intracellular BET BRD Assay kit manual (Promega Corporation, Madison, WI, USA). Briefly cells were transfected and after 24 hours were plated onto flat bottom, non-binding surface, white, polystyrene 96-well plates (Corning Incorporated, Kennebunk, ME, USA). The AGX51 tracer was then added (0–4 μM), followed by digitonin (Sigma, St. Louis, MO, USA) to permeabilize the cells (50 μg/mL). The NanoBRET™ Nano-Glo® Substrate (Promega) was then added and readings taken using a GloMax Discover System instrument (Promega). For the competition assays, cells were treated with 2 μM of AGX51 tracer and 0–60 μM of AGXA or AGX51.
N terminal nanoluc luciferase id1 fusion protein
Manufactured by Genewiz
Sourced in United States
The N-terminal NanoLuc® luciferase ID1 fusion protein is a laboratory tool that combines the NanoLuc® luciferase enzyme with the ID1 protein domain. The NanoLuc® luciferase is a small, bright, and stable luciferase that can be used as a reporter for various biological assays. The ID1 protein domain is a protein involved in cellular processes. This fusion protein allows for the study of the ID1 protein and its interactions within a cellular context.
Automatically generated - may contain errors
Lab products found in correlation
Digitonin, by Merck Group (2 mentions)
Nanobret nano glo substrate, by Promega (2 mentions)
Nanobret te intracellular bet brd assay kit, by Promega (2 mentions)
Glomax discover system instrument, by Promega (2 mentions)
Ecori hf, by New England Biolabs (2 mentions)
Xbai, by New England Biolabs (2 mentions)
2 protocols using n terminal nanoluc luciferase id1 fusion protein
1
NanoBRET Intracellular BET BRD Assay
2
NanoBRET Intracellular BET BRD Assay
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A N-terminal NanoLuc® luciferase ID1 fusion protein was synthesized by Genewiz (South Plainfield, NJ, USA) into the pUC57 backbone and then cloned into the pcDNA3.1 plasmid with EcoRI-HF and XbaI (both enzymes from New England Biolabs, Ipswich, MA, USA). The assay was performed essentially as described in the NanoBRET™ TE Intracellular BET BRD Assay kit manual (Promega Corporation, Madison, WI, USA). Briefly cells were transfected and after 24 hours were plated onto flat bottom, non-binding surface, white, polystyrene 96-well plates (Corning Incorporated, Kennebunk, ME, USA). The AGX51 tracer was then added (0–4 μM), followed by digitonin (Sigma, St. Louis, MO, USA) to permeabilize the cells (50 μg/mL). The NanoBRET™ Nano-Glo® Substrate (Promega) was then added and readings taken using a GloMax Discover System instrument (Promega). For the competition assays, cells were treated with 2 μM of AGX51 tracer and 0–60 μM of AGXA or AGX51.
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