Global RNA Synthesis Assay Kit (Green) (ab273285)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate
- Sample type: Adherent cells, Suspension cells
Overview
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Product name
Global RNA Synthesis Assay Kit (Green)
See all RNA Synthesis kits -
Detection method
Fluorescent -
Sample type
Adherent cells, Suspension cells -
Assay type
Quantitative -
Assay duration
Multiple steps standard assay -
Product overview
Global RNA Synthesis Assay Kit (Green) (ab273285) relies on the incorporation of cell permeable 5-EU (Ethynyl uridine) into nascent RNA, but not into DNA, instead of its natural uridine analog. 5-EU can be used as a replacement for BrU (5-Bromo-uridine) to measure de novo synthesized RNA in proliferating cells. Modified RNA is detected by click chemistry using an azide-containing dye that enables for multiplex analyses with other probes, or detection of RNA-interactive proteins for deeper biological insights. The kit includes Actinomycin D, an inhibitor RNA synthesis that serves as an experimental control.
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Platform
Microplate
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Actinomycin D (100X) 1 x 10µl Copper Reagent (100X) 1 x 100µl Fixative Solution 1 x 10ml Fluorescent Azide (100X) 1 x 100µl Permeabilization Buffer (10X) 1 x 25ml Reducing Agent (20X) 1 x 500µl RNA Label (100X) 1 x 100µl Total DNA Stain (1000X) 1 x 20µl Wash Buffer (10X) 1 x 25ml
Images
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Metabolic labeling of RNA on proliferating HeLa cells
Metabolic labeling of RNA on proliferating HeLa cells. 1 x 105 cells were pre-treated with vehicle or 1 X Actinomycin D for 4h at 37°C prior to 24 hours incubation with RNA Label then processed for detection of de novo synthesized RNA according: Upper panel corresponds to green fluorescence of de novo synthesized RNA. The lower panel shows cells treated with Actinomycin. Nuclear staining confirms that the green signal results from RNA Label incorporation.
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Plate analyses on Jurkat cellsPlate reader analyses on Jurkat cells (1X106 cells/well) with controls and Actinomycin treatment. Average fluorescence standard deviation plotted for 3 replicates per condition.
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Azide Fluorescence Curve in 0-1 X rangeTypical standard curve – data provided for demonstration purposes only. A new standard curve must be generated for each assay performed.
Azide Fluorescence Curve in 0-1 X range. This is reference data and it should not be used to interpret actual results. Your data will depend on the cell type and tested compound.
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Azide Fluorescence Curve of Jurkat cells prepared for this assayTypical standard curve – data provided for demonstration purposes only. A new standard curve must be generated for each assay performed.
Detection limit corresponds to about 31,250 of Jurkat cells per well. A new curve must be obtained for each experiment and the cell line.
