Other (lower) bands are also seen in the blot (see image). We are unsure as to the identity of these - they may be cleavage products of Dcr2, or unspecific bands. These bands are all, however, blocked with the immunising peptide.
RNA interference is an evolutionarily conserved gene silencing pathway in which the endonuclease, Dicer cleaves double stranded RNA into small interfering RNAs.
Dicer is a multidomain protein related to the RNase III protein family. Dicer is required by the RNA interference and small temporal RNA (stRNA) pathways to produce the active small interfering RNA (siRNA) component that represses gene expression. Dicer related RNA interference machinery is also involved in the formation of the heterochromatin structure in organisms such as yeast and higher vertebrate cells. In mammalian cells, both microRNAs (miRNAs) and small interfering RNAs (siRNAs) are thought to be loaded into the same RNA induced silencing complex (RISC), where they guide mRNA degradation or translation silencing depending on the complementarity of the target. Two transcript variants encoding the same protein have been identified.
The Saccharomyces cerevisiae cell cycle regulator genes DCR2 and DCR1 appear to be involved in the initiation of DNA replication. It has been predicted that DCR2 might encode a protein with phosphoesterase activity.
Immunocytochemistry/ Immunofluorescence - Anti-Dcr 2 / Dicer 2 antibody (ab4732)This image is courtesy of an Abreview submitted by Nathalie Gueguen
ab4732 staining Dcr 2 / Dicer 2 (green) in fruit fly (Drosophilia melanogaster) OSS cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.2% Tween and blocked with 1% BSA for 30 minutes at 22°C. Samples were incubated with primary antibody (1/200 in PBS + 0.2% Tween) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. ab24609, anti-Nuclear Pore Complex Proteins, was used as a nuclear membrane marker (red).
This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab4732 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
Harrington AW et al. Drosophila melanogaster retrotransposon and inverted repeat-derived endogenous siRNAs are differentially processed in distinct cellular locations. BMC Genomics18:304 (2017).
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Russo J et al. Antisense Transcription of Retrotransposons in Drosophila: An Origin of Endogenous Small Interfering RNA Precursors. Genetics202:107-21 (2016).
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