Human anti argonaute2 ago2 antibody

The Magna RIP^TM RNA Binding Protein Immunoprecipitation Kit (Millipore, US) was applied for RIP assay to exploring the binding relationship between LINC00482 and miR-2467-3p in PCa cell lines according to the instruction. Cell lysates lysed by RIP lysis buffer were incubated in RIP buffer with magnetic beads conjugated with a negative control IgG (Millipore, US) or human anti-Argonaute 2 (Ago2) antibody (Millipore, US), followed by incubating with proteinase K and precipitation. Purified RNA was used for real time PCR analysis.

Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Billerica, MA, USA) was used for RIP. Cells were lysed in complete RNA lysis buffer; then cell lysates were incubated with RIP buffer containing magnetic beads conjugated with human anti-Argonaute2 (AGO2) antibody (Millipore) or negative control mouse immunoglobulin G (IgG) (Millipore).

The RIP assay was performed using an EZ‐Magna RIP RNA‐binding protein immunoprecipitation kit (Millipor), following the manufacturer's protocol. BMSCs were lysed in RNA lysis buffer containing protease and RNase inhibitors. The protein extract was incubated with RIP wash buffer containing magnetic beads conjugated with a human anti‐argonaute 2 (Ago2) antibody (Millipore) or mouse IgG (Millipore) as a negative control for 2 hr at 4 °C. Unbound proteins were digested with proteinase K, and co‐immunoprecipitated RNA was isolated. Then, qRT‐PCR analysis was performed to determine the presence of binding targets.

The EZMagna RNA immunoprecipitation (RIP) Kit (Millipore, USA) was used according to the manufacturer’s protocol. HNE1 and C666-1 cells were lysed and incubated with RIPA buffer containing magnetic beads conjugated with a human anti-Argonaute2 (Ago2) antibody (Millipore). Normal mouse IgG (Millipore) was used as a negative control. The samples were incubated with proteinase K, after which immunoprecipitated RNA was extracted. Purified RNA was subjected to qRT-PCR analysis.

Magna RIP™ RNA-Binding Protein Immunoprecipitation kit (Millipore, Billerica, MA, USA) were used for RIP. NSCLC cells were lysed in complete RNA lysis buffer, then cell lysates were incubated with RIP immunoprecipitation buffer containing magnetic beads conjugated with human anti-argonaute 2 (AGO2) antibody (Millipore) or negative control mouse IgG (Millipore). Extracted RNAs were analyzed by RT-qPCR to identify the presence of circRNA-FOXO3.
The RIP experiment using FOXO3 antibody (1:1,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) to pull down miR-155 was also performed. The RIP RNA fraction was digested by DNase and cDNA was generated using PrimeScript 1st strand cDNA Synthesis kit (Takara Bio., Inc., Otsu, Shiga, Japan). Final analysis was investigated by performing RT-qPCR and presented as a fold enrichment of miR-155.

The RNA immunoprecipitation (RIP) assay was carried out according to the manufacturer's instructions using the Magna RIP^TM RNA‐binding protein immunoprecipitation kit (Millipore). In brief, cells were lysed in RIP lysis buffer and the cell lysates were incubated with RIP buffer containing magnetic beads conjugated with human anti‐Argonaute2 (AGO2) antibody (Millipore) or negative control mouse IgG (Millipore). The magnetic beads coated with antibodies were added to the extracted RNA suspension and incubated on ice for 30 min. After eluting the magnetic beads, we extracted the RNA and purified by using the purification kit. Then, PCR was used for quantitative analysis.