Ataxin-2-HiBiT cells were lysed in-well with 125μL of Nano-Glo® HiBiT Lytic Buffer, and lysate from one well was split and used for detection of both HiBiT- and FFLuc-generated luminescence. 25μL of lysate was placed in two wells of an opaque white, flat bottom 96-well assay plate (Sigma Aldrich, cat# CLS3990). For HiBiT detection, 25μL of HiBiT lytic reagent (1:25 substrate, 1:50 LgBiT) was added to the lysate (Promega, cat# N3050). For FFluc detection, 25μL of ONE-Glo Assay Buffer (Promega, cat# E6120) was added to the lysate. Plates were incubated in the dark with gentle rotation for 10 min, then luminescence was measured on a Tecan Spark plate reader (Tecan). For all assays, HiBiT signal was normalized to the FFluc signal for each individual well of the original transfected plate then normalized to the non-targeting/untreated control, this value is represented in bar graphs.
One glo assay buffer
Manufactured by Promega
The ONE-Glo Assay Buffer is a laboratory reagent designed to measure cellular ATP levels. It is a key component of the ONE-Glo Luciferase Assay System, which is used to quantify ATP as an indicator of metabolic activity or cell viability.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using one glo assay buffer
1
Dual Luminescence Assay for Ataxin-2
2
Dual Luminescence Assay for Ataxin-2
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Ataxin-2-HiBiT cells were lysed in-well with 125µL of Nano-Glo® HiBiT Lytic Buffer, and lysate from one well was split and used for detection of both HiBiT-and FFLuc-generated luminescence. 25µL of lysate was placed in two wells of an opaque white, flat bottom 96-well assay plate (Sigma Aldrich, cat# CLS3990). For HiBiT detection, 25µL of HiBiT lytic reagent (1:25 substrate, 1:50 LgBiT) was added to the lysate (Promega, cat# N3050). For FFluc detection, 25µL of ONE-Glo Assay Buffer (Promega, cat# E6120) was added to the lysate. Plates were incubated in the dark with gentle rotation for 10 minutes, then luminescence was measured on a Tecan Spark plate reader (Tecan). For all assays, HiBiT signal was normalized to the FFluc signal for each individual well of the original transfected plate then normalized to the non-targeting/untreated control, this value is represented in bar graphs.
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