RNA immunoprecipitation (RIP) was done as described elsewhere [21 (link)]. Determination of the RIP was done with a Magna RIP Kit (Millipore, Billerica, MA, USA). To this end, collection and resuspension of 1 × 107 OS cells were performed in 100 μL of RIP lysis buffer plus protease inhibitor cocktail and RNase inhibitors. Approximately 200 μL of the cell lysates were then incubated with 5 μg magnetic beads conjugated to anti-Argonaute 2 (AGO2) antibody (Millipore) or control rabbit IgG (Millipore). After digestion with proteinase K, extraction of the immunoprecipitated RNAs was performed. Test of enrichment of circ_ANKIB1and miR-26b-5p was finally conducted. The experiment was done in triplicate.
Anti argonaute2 ago2 antibody
Manufactured by Merck Group
Sourced in United States
The Anti-Argonaute2 (Ago2) antibody is a research tool used to detect and study the Ago2 protein, a key component of the RNA-induced silencing complex (RISC). Ago2 plays a crucial role in gene expression regulation through RNA interference (RNAi) pathways. This antibody can be used in various applications, such as Western blotting, immunoprecipitation, and immunocytochemistry, to identify and analyze the Ago2 protein in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Magna rip rna binding protein immunoprecipitation kit, by Merck Group (9 mentions)
Normal mouse igg, by Merck Group (5 mentions)
Control rabbit igg, by Merck Group (2 mentions)
Magna rna binding protein immunoprecipitation kit, by Merck Group (2 mentions)
Magna rip kit, by Merck Group (1 mentions)
Control igg, by Merck Group (1 mentions)
Normal mouse igg antibody, by Merck Group (1 mentions)
Ez magna rip kit, by Merck Group (1 mentions)
Rip lysis buffer, by Merck Group (1 mentions)
Magna riptm rna binding protein immunoprecipitation kit, by Merck Group (1 mentions)
Ez magna rip rna binding protein immunoprecipitation kit, by Merck Group (1 mentions)
Qiaamp minelut virus spin kit, by Qiagen (1 mentions)
17 protocols using anti argonaute2 ago2 antibody
1
AGO2-Bound CircRNA and miRNA Detection
2
LAMTOR5-AS1 Interactome Profiling
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By utilizing a Magna RIP RNA Binding Protein Immunoprecipitation Kit (Merck Millipore, Darmstadt, Germany), we examined the interaction among LAMTOR5-AS1, miR-506-3p, and E2F6. In brief, whole NSCLC cell lysates were obtained by treatment of cells with complete RIP lysis buffer and probed overnight at 4°C with RIP buffer supplemented with magnetic beads containing anti-Argonaute2 (Ago2) antibody or IgG control (Millipore). Then the magnetic beads were harvested and rinsed using RIP washing buffer, followed by treatment with Proteinase K buffer. The target immunoprecipitated RNA was extracted and detected via qRT-PCR.
3
Argonaute 2-mediated RNA Immunoprecipitation
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RIP assay was conducted using the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Billerica, MA, USA). OS cells were lysed in complete RIP lysis buffer and the cell lysate was incubated with RIP buffer containing magnetic beads conjugated with an anti-Argonaute 2 (Ago2) antibody or normal mouse IgG antibody (Millipore, Bedford, MA, USA). Following an overnight incubation at 4°C, the magnetic beads were harvested and rinsed with wash buffer. The resulting immunoprecipitate complex was incubated with proteinase K to purify RNA. Finally, the relative enrichment of LINC00839 and miR-454-3p in the immunoprecipitated RNA was determined by qRT-PCR.
4
Validation of circ_ZNF124-miR-337-3p Interaction
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RNA immunoprecipitation used to verify circ_ZNF124 and miR-337-3p interaction in vivo was performed by using Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Bedford, MA). In brief, 20 million A549 cells were harvested and lysed with RIP lysis buffer containing protease and RNase inhibitors. Cell lysates were then split, and RNA was pulled down by incubating with anti-Argonaute 2 (AGO2) antibody (Millipore) or control rabbit IgG (Millipore), followed by rotating at 4 °C overnight. After proteinase K treatment, the immunoprecipitated RNAs were extracted. The abundance of circ_ZNF124 and miR-337-3p were determined by qRT-PCR.
5
Argonaute 2 RNA Immunoprecipitation
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RNA immunoprecipitation (RIP) assay was performed using the EZMagna RIP-Kit (Millipore). Target cells were lysed in RIP lysis buffer and then conjugated to anti-Argonaute 2 (AGO2) antibody (Millipore) or control anti-immunoglobulin G (IgG) antibody (Millipore) for two hours at 4°C. Purified RNA was analyzed using qRT-PCR after beads were washed with RIP buffer.
6
RIP of SBF2-AS1 and miR-338-3p
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RIP was performed using a Magna RNA-binding protein immunoprecipitation kit (Millipore, Billerica, MA, USA) following the manufacturer's instructions. U87 and U118 cell lysates containing SBF2-AS1 and miR-338-3p were prepared and incubated with anti-argonaute2 (Ago2) antibody (Millipore). Normal mouse IgG (Millipore) was used as a NC. SBF2-AS1 and miR-338-3p present in the precipitates were assayed by qRT-PCR.
7
Profiling of miRNA-Argonaute Interactions
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TPC-1 cells (~1 × 107) transfected with miR-98 mimic or mimic-NC for 48 h were washed with cold PBS and lysed with RNA immunoprecipitation (RIP) lysis buffer (EMD Millipore, Billerica, MA, USA). Cell lysates were incubated with magnetic beads conjugated with anti-Argonaute2 (AGO2) antibody (Millipore) or mouse IgG that served as a negative control (Millipore). Beads were collected and washed, and RNA was extracted in the presence of proteinase K. OIP5-AS1 was detected by RT-PCR and qRT-PCR. The cell lysate served as the input.
8
RIP Assay for AGO2-bound RNAs
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RIP assays were carried out in human BMECs after utilizing the Magna RIP RNA-binding protein immunoprecipitation kit (Millipore, Billerica, MA, USA). Cells (1×107) were lysed in complete RNA lysis buffer, then cell lysates were incubated with anti-Argonaute2 (AGO2) antibody (Millipore, Billerica, Ma, USA), negative control mouse IgG (Millipore, Billerica, Ma, USA) or total RNAs (input controls) in accordance with the manufacturer’s instructions. Co-precipitated RNA was measured by RT-qPCR as previously described.
9
RIP Assay for miRNA Targets
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RIP assays were carried out in SH-SY5Y cells pretreated with MPP+, 48 hours after transfection with miR-1277-5p or miR-NC, utilizing the Magna RIP RNA-binding protein immunoprecipitation kit (Millipore, Billerica, MA, USA). Cells (1 × 107) were lysed in complete RNA lysis buffer, then cell lysates were incubated with anti-Argonaute 2 (AGO2) antibody (Millipore, Billerica, Ma, USA), negative control mouse IgG (Millipore, Billerica, Ma, USA) or total RNAs (input controls) in accordance with the manufacturer's instructions. Coprecipitated RNA was measured by qRT-PCR as previously described.
10
NEAT1-miR-129 Interaction Analysis
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To verify the relationship between NEAT1 and miR-129, RIP was conducted using the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Millipore). Briefly, EC109 and EC9706 cells at 80% confluency were harvested and lysed in complete RIP lysis buffer. Afterwards, the whole cell extract (100 μl) was coimmunoprecipitated with RIP buffer containing magnetic beads conjugated with anti-Argonaute2 (Ago2) antibody (Millipore) or normal mouse IgG (Millipore) as a negative control. Then, samples were digested with proteinase K, and then, coprecipitated RNA was isolated and subjected to qRT-PCR analysis of NEAT1 and miR-129.
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