Human anti argonaute2 anti ago2 antibody

To verify the direct interaction between PVT1 and miR-16, RIP assay was performed using an EZ-Magna RIP Kit (Millipore, Billerica, MA, USA) according to the manufacturer’s instruction. NCI-N87 and MKN45 cells were lysed using RIP lysis buffer containing protease inhibitor and RNase inhibitor, followed by the incubation with RIP buffer containing magnetic beads conjugated with human anti-argonaute 2 (anti-Ago2) antibody (Millipore, Billerica, MA, USA) or negative control normal mouse IgG (Millipore, Billerica, MA, USA) for 1 h. Subsequently, after protein removal with proteinase K, the immunoprecipitated RNA was extracted for qRT-PCR analysis.

RIP™ RNA-Binding Protein Immunoprecipitation Kit (Millipore, Billerica, MA, USA) was used for RIP. GBM cell lysates were prepared and incubated with RIP buffer containing magnetic beads conjugated with human anti-Argonaute2 (anti-Ago2) antibody (Millipore, Billerica, MA, USA) while normal mouse IgG (Millipore, Billerica, MA, USA) functioned as a negative control. Immunoprecipitated RNA was tested by qRT-PCR analysis to verify the presence of lncSBF2-AS1 using specific primers.

RIP assay was performed using the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore) in agreement with the manufacturer’s instruction. SKOV3 and OVCAR3 cell lysates were obtained and incubated with RIP buffer containing magnetic beads conjugated with human anti-Argonaute2 (anti-Ago2) antibody (Millipore) or normal mouse IgG (control; Millipore). RNA was extracted from immunoprecipitate and analyzed by qRT-PCR.