Dicer, also known as endoribonuclease Dicer or helicase with RNase motif, is an enzyme that in humans is encoded by the DICER1 gene. Being part of the RNase III family, Dicer cleaves double-stranded RNA (dsRNA) and pre-microRNA (pre-miRNA) into short double-stranded RNA fragments called small interfering RNA and microRNA, respectively. These fragments are approximately 20–25 base pairs long with a two-base overhang on the 3′-end. Dicer facilitates the activation of the RNA-induced silencing complex (RISC), which is essential for RNA interference. RISC has a catalytic component Argonaute, which is an endonuclease capable of degrading messenger RNA (mRNA).
Background References
1. Poirier EZ et al. An isoform of Dicer protects mammalian stem cells against multiple RNA viruses. Science. 2021 Jul
2. Wang Q et al. Mechanism of siRNA production by a plant Dicer-RNA complex in dicing-competent conformation. Science. 2021 Nov
Western blot analysis of Dicer on different lysates with Rabbit anti-Dicer antibody (HA500522) at 1/5,000 dilution.
Lane 1: HEK-293 (Human embryonic kidney cell) cell lysate Lane 2: Daudi (Human Burkitt's lymphoma cell) cell lysate
Lysates/proteins at 15 µg/Lane. Exposure time: 14 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA500522, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 218 kDa Observed band size: 250 kDa
Immunocytochemistry analysis of MCF-7 cells labeling Dicer with Rabbit anti-Dicer antibody (HA500522) at 10ug/mL dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Dicer antibody (HA500522) at 10ug/mL dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Flow cytometric analysis of C2C12 cells labeling Dicer.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA500522, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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