Dicer Antibody

Western Blot: Dicer Antibody [NBP1-06520]

Western Blot: Dicer Antibody [NBP1-06520] - Whole cell lysate (15 ug) from HeLa, 293T, Jurkat, mouse TCMK-1, and mouse NIH3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit antiDicer antibody used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.

Immunoprecipitation: Dicer Antibody [NBP1-06520]

Immunoprecipitation: Dicer Antibody [NBP1-06520] - Detection of human Dicer by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HEK293T cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-Dicer antibody NBP1-06520 (lot NBP1-06520-5) used for IP at 6 ug per reaction. Dicer was also immunoprecipitated by a previous lot of this antibody (lot NBP1-06520-4). For blotting immunoprecipitated Dicer, NBP1-06520 was used at 1 ug/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.

Western Blot: Dicer Antibody [NBP1-06520] -

Western Blot: Dicer Antibody [NBP1-06520] - Nef does not affect siRNA loading into RISC, but inhibits its slicing activity.(A) Lysates from the U937 stable cell lines were prepared in Buffer D as described in Materials & Methods. From this, 25 µg of lysate was incubated with 32P-labeled duplex siRNA (without or with 50-fold molar excess of unlabeled siRNA) in binding buffer for 30 min at 4°C. The complexes were separated on a 6% non-denaturing polyacrylamide gel. (B) The gel shift assay was set up as in (A) except that lysates immunodepleted for either Ago2 or Dicer were also included. The positions of the mobility shifted complexes & free probe are indicated. (C) Slicer activity was assayed using immunoprecipitated Ago2 from U937/Nef-EYFP or U937/EYFP cell lysates & a 32P-labeled in vitro transcribed let7a RNA, as described in Materials & Methods. The positions of full-length RNA & the two sliced products are shown. The 59 nt marker oligonucleotide was run on the same gel, but a lower exposure is shown. (D) Densitometric analysis of autoradiograms from three independent experiments was carried out using Image J (version 1.4.1). The slicing activity in Nef-EYFP lysates is represented as a percentage of that in EYFP lysates. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0074472), licensed under a CC-BY license. Not internally tested by Novus Biologicals.