Agonist-mediated internalization of ACKR3 was measured by BRET2 between ACKR3_RlucII and rGFP_CAAX (a gift from M. Bouvier, Université de Montréal) as previously described24 (link). Samples were prepared as described for β-arrestin2 recruitment time courses with the exception of the transfected DNA amounts. HEK293 cells were transfected with 42 ng ACKR3_RlucII and 170 ng rGFP_CAAX DNA, with empty pcDNA3.1 to bring the total DNA amount to 2.5 µg/well. Data presented in Figs. 1 , 3 , and 4C were collected with a PerkinElmer Victor Luminometer, while the time courses in Fig. 4D and 7 were measured on a Tecan Spark luminometer. All settings were identical to those used for arrestin association (described above). Data is presented as percent change compared to mock treated wells and are a composite of three independent experiments. The percent changes after 30 min were compared for statistical analysis.
Victor luminometer
Manufactured by Tecan
The Victor Luminometer is a compact and versatile instrument designed for luminescence-based detection and quantification. It employs a sensitive photomultiplier tube to accurately measure light signals generated by a variety of sample types, including cell-based assays, biochemical assays, and reporter gene assays.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using victor luminometer
1
ACKR3 Internalization Measurement by BRET2
2
ACKR3 Internalization Measured by BRET2
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Agonist-mediated internalization of ACKR3 was measured by BRET2 between ACKR3_RlucII and rGFP_CAAX (a gift from M. Bouvier, Université de Montréal) as previously described (Namkung et al., 2016 (link)). Samples were prepared as described for β-arrestin2 recruitment time courses, with the exception of the transfected DNA amounts. HEK293 cells were transfected with 42 ng ACKR3_RlucII and 170 ng rGFP_CAAX DNA, with empty pcDNA3.1 to bring the total DNA amount to 2.5 µg/well. Data presented in Figs. 1 , 3 , and 4C were collected with a PerkinElmer Victor Luminometer, while the time courses in Figs. 4D and 7 were measured on a Tecan Spark luminometer. All settings were identical to those used for arrestin association (previously described). Data are presented as percent change compared to mock-treated wells and are a composite of three independent experiments. The percent changes after 30 minutes were compared for statistical analysis.
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