Clone OTI4C5, Anti-DDK (FLAG) Monoclonal Antibody, Agarose Beads

Snippet: ransfection Transfection of ISK cells was performed twenty-four hours after seeding cells on 6 well plates at a density of 0.5 × 106 cells/well by using a mixture of MYC-DDK tagged Dyskerin plasmid (OriGene Technologies, Rockville, MD, USA, 3 μL) with Lipofectamine 2000 (Thermo Fisher Scientific, Loughborough, UK, 9 μL). The plasmid or the Lipofectamine was diluted in 250 μL of Gibco Opti-MEM

Snippet: HEPHL1 Δexon 5) alleles. These were created by site-directed mutagenesis using a commercially available HEPHL1 expression vector with a C-terminal myc-DDK tag (Origene, Rockville, MD) as template. Transfection of these Cached;

Snippet: All rights reserved. siRNA (Origene, Rockville, MD, USA) diluted in silentFect reagent (BioRad). Control group After 72 hours of transfection with either scrambled siRNA control (Santa Cruz) or Kvβ2 siRNA (Origene), BrdU solution (10µM) was added to cells

Snippet: For immuno-precipitation (IP), supernatants obtained from clarified cell lysates were incubated with anti-FLAG agarose beads (Origene) for 12h at 4oC

Snippet: Recombinant FLAG-TRF2, myc-TRF2 and myc-PRL-3 (all from OriGene) were expressed in human HEK293 cells and purified