Lipofectamine™ MessengerMAX™ (Life Technologies) was diluted in opti-MEM to the desired working concentration and dispensed onto 384-well white assay plates (Greiner 781080). A source plate (Labcyte LP-0200) containing serial dilutions of the mRNAs was prepared using the Bravo liquid handler (Agilent), and a 10-point 2-fold dose titration of each mRNA was dispensed onto the assay plate using Echo555 (Labcyte). After a 10 min incubation, 4000 MIA PaCa-2 or SJCRH30 cells or 6000 BJ cells were added per well. For kinetic monitoring, 20 μM Endurazine (Promega), an extended time-released live cell substrate, was added to each well. Luminescence was measured continuously at 1 h intervals for 48 h on the Tecan Spark 10M set to 37ºC, 5% CO2. For end-point HiBiT protein detection, the NanoGlo HiBiT lytic detection assay (Promega, N3040) was performed as per the manufacturer's instructions. Luminescence signal was determined using the Envision plate reader, and values were normalized to a HiBiT-control protein (Promega, N3010)
N3010
Manufactured by Promega
The N3010 is a laboratory product offered by Promega. It is a device designed for specific laboratory functions, but a detailed description while maintaining an unbiased and factual approach is not available.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using n3010
1
Transient mRNA Transfection Assay
2
Transient mRNA Transfection Assay
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Lipofectamine™ MessengerMAX™ (Life Technologies) was diluted in opti-MEM to the desired working concentration and dispensed onto 384-well white assay plates (Greiner 781080). A source plate (Labcyte LP-0200) containing serial dilutions of the mRNAs was prepared using the Bravo liquid handler (Agilent), and a 10-point 2-fold dose titration of each mRNA was dispensed onto the assay plate using Echo555 (Labcyte). After a 10 min incubation, 4000 MIA PaCa-2 or SJCRH30 cells or 6000 BJ cells were added per well. For kinetic monitoring, 20 μM Endurazine (Promega), an extended time-released live cell substrate, was added to each well. Luminescence was measured continuously at 1 h intervals for 48 h on the Tecan Spark 10M set to 37ºC, 5% CO2. For end-point HiBiT protein detection, the NanoGlo HiBiT lytic detection assay (Promega, N3040) was performed as per the manufacturer's instructions. Luminescence signal was determined using the Envision plate reader, and values were normalized to a HiBiT-control protein (Promega, N3010)
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