Human ago2 antibody

OSCC cell lysates were incubated with RIP buffer with magnetic beads conjugated with normal rabbit IgG (negative control) or human Ago2 antibody (Millipore, Bedford, MA). To isolate the immunoprecipitated RNA, the protein was digested by proteinase K. Finally, purified RNAs were analyzed by qRT-PCR.

An RIP assay was performed using the Magna RIP RNA-Binding Protein Immunoprecipitation kit (Millipore Sigma, Burlington, MA, USA). Breast cancer cells were collected and lysed in RIP buffer, and the whole-cell lysates obtained were incubated with RIP buffer containing magnetic beads conjugated with human Ago2 antibody (Millipore). Normal immunoglobulin (Ig) G (Millipore) served as the NC. The magnetic beads were collected after overnight incubation. The immunoprecipitated RNA was extracted and analyzed using RT-qPCR to assess whether LINC02163 and miR-511-3p were coimmunoprecipitated.

RIP assays were carried out to determine the interaction between ZFAS1 and miR-590-3p as in the previous study^{19 (link),20} . We used EZMagna RIP RNA-binding protein immunoprecipitation kit (Millipore, Chengdu, Sichuan, China) according to the manufacturer’s protocol. Cells were subjected to different treatments and then lysed using RNA lysis buffer with 1X protease inhibitor cocktail. The cell lysate was incubated with magnetic beads conjugated with human Ago2 antibody (Millipore, Chengdu, Sichuan, China) or negative control immunoglobulin G (IgG) (Abcam, Beijing, China) to immunoprecipitate the RNA-induced silencing complex. After overnight incubation at 4°C, the coprecipitated RNAs were reverse-transcribed and analyzed by qRT-PCR.