Dicer Fluorogenic Assay Kit

• Background:

  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.

• Description:

  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.Need us to run inhibitor screens or profile your compounds against Dicer? Check out ourMetabolic Enzyme Screening Services.

• UniProt:

  Q9UPY3

• Applications:

  Screen molecules that inhibit Dicer RNAse activity in drug discovery High-Throughput Screening (HTS) applications.Determine compound IC50.Perform Dicer real-time kinetics.

• Format:

  Catalog #NameAmountStorage79083Dicer, His-Tag (Human) *45 µg-80°CPre-micro-RNA Substrate 15 µl-80°CDR-01 Buffer3 ml-20°C384-well black microplate1Room Temp* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.

• Shipping Conditions:

  -80°C

• Storage Conditions:

  This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

• Notes:

  Troubleshooting GuideVisitbpsbioscience.com/assay-kits-faqfor detailed troubleshooting instructions. For further questions, please email[email protected]