Pierce magnetic rna protein pull down

HAR1A RAN pull-down assay was conducted using the Pierce™ Magnetic RNA-Protein Pull-Down (Thermo Scientific, USA) and MAXIscript® (Ambion, USA) kits based on protocols provided by manufacturers. Cells lysed in RIPA buffer were incubated with in vitro transcribed full-length HAR1A probes and antisense probes labeled by biotins (Gene Create, China). Briefly, NSCLC cell extracts were incubated with biotinylated HAR1A or antisense-HAR1A (control), affinity precipitated using streptavidin-conjugated beads. The RNA-binding protein complexes were collected, washed, eluted, and analyzed by Liquid Chromatography and Mass Spectrometry (LC/MS) and western blot.

The Pierce™ Magnetic RNA-Protein Pull-Down (Thermo Scientific, USA) was used for this experiment. Cell extracts were mixed with biotinylated KCNK15-AS1 sense or KCNK15-AS1 antisense and streptavidin beads. After washing and boiling, the RNA complex was detected via western blot. The experiments were conducted three times.

The Pierce™ Magnetic RNA-Protein Pull-Down (Thermo Scientific, USA) and MAXIscript® (Ambion, USA) kits were used for this assay based on provided protocols. Briefly, PANC-1 nuclear extracts were combined with biotinylated CF129 or antisense- CF129, after which streptavidin beads were added to the samples. Pulled down RNA was then isolated by washing these beads, boiling them in SDS buffer, and subsequent western immunoblotting.