Anti dicer

Manufactured by Merck Group

Sourced in United States

Anti-Dicer is a laboratory equipment product designed to inhibit the activity of the Dicer enzyme. Dicer is an enzyme involved in the processing of double-stranded RNA (dsRNA) into short interfering RNA (siRNA) and microRNA (miRNA). Anti-Dicer can be used to study the role of Dicer in various cellular processes.

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3 protocols using anti dicer

Western Blot Analysis of Cellular Markers

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Cell lysates were prepared with Laemmli buffer supplemented with protease inhibitors (complete tablet; Roche) and phosphatase inhibitors (PhosSTOP tablet; Roche) and analysed by Western blotting with the following primary antibodies: anti-γH2AX (Abcam); anti-PTB (Abcam); anti-nPTB (Abcam); anti-βIII Tubulin (Abcam); anti-Tyrosine Hydroxylase (Abcam); anti-GAPDH (Thermo Scientific); anti-MAP2 (Novusbio); anti-DROSHA (Cell Signaling); anti-DICER (Sigma); anti-H2AX (Millipore). Primary antibodies were revealed with peroxidase-conjugated goat anti-mouse or anti-rabbit antibodies (Jackson Immunoresearch Laboratories) and enhanced chemiluminescence system (Super Signal West Pico Pierce or Super Signal West Dura Extended). βIII Tubulin antibody was revealed with alkaline phosphatase-conjugated goat anti-chicken antibody (Santa Cruz) with a colorimetric assay.

Pignataro D., Francia S., Zanetta F., Brenna G., Brandini S., Olivieri A., Torroni A., Biamonti G, & Montecucco A. (2017). A missense MT-ND5 mutation in differentiated Parkinson Disease cytoplasmic hybrid induces ROS-dependent DNA Damage Response amplified by DROSHA. Scientific Reports, 7, 9528.

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Characterization of Anti-Ago1x Antibody

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Polyclonal anti-Ago1x antibody was generated in rabbit against the unique peptide sequence, RQNAVTSLDRRKLSKP, and was affinity-purified (Pierce Biotechnology). To confirm the specificity, the anti-Ago1x antibody was incubated with the same peptide at a final concentration of 1 μg/ml for 2 h with gentle agitation at room temperature before using it for Western blot (described below). Anti-Ago1 antibody (Cell Signaling Technologies, 9388; Sigma, SAB4200065), anti-Dicer (Sigma, SAB4200087), anti-HA (Sigma, clone 3F10, 11867423001; and Cell Signaling Technology, clone 6E2, 2367), anti-GW182 (Bethyl Laboratories, A302-329A), anti-puromycin (Merck Millipore, clone 12D10, MABE343), anti-Myc (Cell Signaling Technologies, 22725), anti-FLAG (Sigma, clone M2, F1804), anti-Actin (Sigma, A3854), anti-GAPDH (Sigma, G9295), and stabilized peroxidase-conjugated secondary antibodies (Thermo Fisher Scientific) were used at the manufacturer-recommended dilutions. Alexa Fluor-conjugated secondary antibodies (Molecular Probes) were used for immunofluorescence.

Singh A., Manjunath L.E., Kundu P., Sahoo S., Das A., Suma H.R., Fox P.L, & Eswarappa S.M. (2019). Let‐7a‐regulated translational readthrough of mammalian AGO1 generates a microRNA pathway inhibitor. The EMBO Journal, 38(16), e100727.

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Western Blotting Technique for Protein Analysis

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For western blots, samples were separated on SDS-polyacrylamide gel electrophoresis gels and then transferred to Polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). Membranes were processed following the ECL western blotting protocol (GE Healthcare). These antibodies were used in western blots: anti-CENPC1 (Abcam, 1:1 000 dilution, cat no. ab50974), anti-Phospho-Histone H3 (Cell Signaling Technology, 1:1 000 dilution, cat no. 9706), anti-ACTB (Abcam, 1:1 000 dilution, cat no. ab8227), anti-Dicer (Sigma, 1:2 000 dilution, cat no. SAB4200087), anti-GAPDH (Cell Signaling Technology, Danvers, MA, USA, 1:2 000 dilution, cat no. 3683), anti-AGO2 (Sigma, 1:2 000 dilution, cat no. SAB4200085) and anti-HDAC2 (Cell Signaling Technology, 1:1 000 dilution, cat no. 5113). Images were taken with LAS-4000mini Image Reader (GE Healthcare).

Huang C., Wang X., Liu X., Cao S, & Shan G. (2015). RNAi pathway participates in chromosome segregation in mammalian cells. Cell Discovery, 1, 15029-.

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