RIP was performed using a Magna RIP RNA-Binding Protein Immunoprecipitation kit (Millipore, Bedford, MA) according to the manufacturer’s instructions. Briefly, 2 × 107 NOZ cell lysates were incubated with magnetic beads conjugated with negative control normal mouse IgG or human anti-Ago2 antibody (Millipore). The immunoprecipitated RNAs were then extracted and detected by qRT-PCR to confirm the enrichment of binding targets and the products were then subjected to agrose gel electrophoresis. The primers used for detecting lncRNA-HGBC or miR-502-3p were listed in Additional file 1 : Table S2.
Normal mouse igg or human anti ago2 antibody
Manufactured by Merck Group
Normal mouse IgG is an immunoglobulin G (IgG) antibody purified from normal mouse serum. It is commonly used as a control antibody in various immunological assays and experiments. Human anti-Ago2 antibody is an antibody that specifically binds to the Argonaute 2 (Ago2) protein, which is a key component of the RNA-induced silencing complex (RISC) involved in gene silencing mechanisms.
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Lab products found in correlation
4 protocols using normal mouse igg or human anti ago2 antibody
1
Identification of lncRNA-HGBC and miR-502-3p
2
Investigating miR-15a-5p and CERS6-AS1 Interactions
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The possible interactions between miR-15a-5p and CERS6-AS1 were examined with a Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Bedford, MA, USA). RIP lysis buffer was utilized to lyse PDAC cells, after which 10 μl of the cell lysate was retained as input and 100 μl of the cell lysate were probed overnight at 4° C using magnetic beads conjugated with either normal mouse IgG or human anti-Ago2 antibody (Millipore) suspended in 900 μl of RIP lysis buffer. Magnetic beads were then collected and treated with Proteinase K to digest the proteins. The immunoprecipitated RNA was evaluated via RT-qPCR to measure CERS6-AS1 and miR-15a-5p enrichment.
3
Enrichment of UBA6-AS1 and miR-760 in GBM cells
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A Magna RIP RNA-Binding Protein Immunoprecipitation Kit (EMD Millipore) was applied in RIP assay. Briefly, GBM cell suspension was obtained by incubation with RIP buffer (Beyotime Institute of Biotechnology). Subsequently, the cell suspension was cultured with magnetic beads pre-incubated with human anti-Ago2 antibody or normal mouse IgG (EMD Millipore). Following overnight incubation at 4°C, the magnetic beads were harvested and treated with Proteinase K for protein digestion. The precipitated RNA was extracted and analyzed by RT-qPCR to detect the enrichment of UBA6-AS1 and miR-760.
4
RNA Binding Protein Immunoprecipitation Assay
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RIP assay was performed to test the binding interaction among LINC01124, miR-1247-5p, and FOXO3 via RNA-induced silencing complexes (RISCs) using the Magna RIP RNA Binding Protein Immunoprecipitation Kit (Merck Millipore, Darmstadt, Germany). Briefly, HCC cell extracts were prepared using RIP lysis buffer, followed by incubation with magnetic beads conjugated with human anti-Ago2 antibody or normal mouse IgG (Merck Millipore). Subsequently, the magnetic beads were collected and incubated with Proteinase K to remove any proteins. The immunoprecipitated RNA was measured by qRT-PCR. IgG acted as a negative control, while input functioned as the positive control.
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