Dicer Fluorogenic Assay Kit
The Dicer Fluorogenic Assay Kit is a homogeneous 384-well assay designed to measure the double-stranded-RNA processing activity of Dicer for screening and profiling applications. The Dicer Fluorogenic Assay Kit contains enough purified Dicer, substrate, and assay buffer for 384 reactions.
Figure 1: Illustration of the Dicer Fluorogenic Assay method.
The assay relies on an RNA duplex imitating pre-miRNA, containing a conjugated fluorophore and quencher pair. Fluorescence of the dye is effectively quenched by the proximity of the quencher in the intact RNA duplex. The endoribonuclease Dicer processes the pre-miRNA into a mature 22 bp double-stranded miRNA by cleaving the 5’ end which is conjugated to the dye, releasing it from the miRNA. Separated from the quencher, the dye emits fluorescence with excitation/emission maxima of λ=647/ 665 nm. Dicer activity is proportional to the increase in fluorescence.
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- Fluorescent microplate reader capable of measuring excitation/emission maxima of λ=647/ 665 nm
- Adjustable micropipettor and sterile tips
- Rotating or rocker platform
| Catalog # | Name | Amount | Storage |
| 79083 | Dicer, His-Tag (Human)* | 45 µg | -80°C |
| Pre-micro-RNA Substrate 1 | 5 µl | -80°C | |
| DR-01 Buffer | 3 ml | -20°C | |
| 384-well black microplate | 1 | Room Temp |
* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.
Dicer (also known as endoribonuclease Dicer or helicase with RNase motif) is a ribonuclease that participates in pre-microRNA (pre-miRNA) and double stranded RNA (dsRNA) cleavage. The cleavage of pre-miRNA into mature miRNA and formation of small interfering RNA (siRNA) from dsRNA are essential for gene regulation and DNA repair. Decreased levels of Dicer can result in Age-related Macular Degeneration (AMD), the most common age-related blindness. Dicer impairment is also involved in cancer and RNA-based viral infection. The development of small molecule inhibitors or other strategies that affect Dicer activity can provide new therapeutical avenues for cancer treatment and AMD prevention.
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