According to the instructions of RIP kit (Millipore, Bedford, MA, USA), Daoy and D341 cells were lysed in RIP lysate. The magnetic beads were preincubated with antibody IgG (Millipore) and anti-Ago2 (Millipore) at 37°C for 1 h. The cell lysates were immunoprecipitated by the immunoprecipitation method and overnight at 4°C, then the detection of purified immunoprecipitated RNA by qRT-PCR.
Anti ago2
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom
Anti-Ago2 is a laboratory reagent that specifically binds to and inhibits the activity of the Argonaute 2 (Ago2) protein. Ago2 is a key component of the RNA-induced silencing complex (RISC), which plays a central role in RNA interference (RNAi) pathways. Anti-Ago2 can be used to study the function of Ago2 and its involvement in gene silencing mechanisms.
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Lab products found in correlation
Magna rip rna binding protein immunoprecipitation kit, by Merck Group (21 mentions)
Ez magna rip kit, by Merck Group (8 mentions)
Magna rip kit, by Merck Group (8 mentions)
Control igg, by Merck Group (8 mentions)
Imprint rna immunoprecipitation kit, by Merck Group (5 mentions)
Rip lysis buffer kit, by Merck Group (5 mentions)
Magna riptm rna binding protein immunoprecipitation kit, by Merck Group (4 mentions)
Rip kit, by Merck Group (3 mentions)
Magnetic bead, by Thermo Fisher Scientific (3 mentions)
Rip buffer, by Merck Group (3 mentions)
Ez magna rip rna binding protein immunoprecipitation kit, by Merck Group (3 mentions)
Magna rna binding protein immunoprecipitation kit, by Merck Group (3 mentions)
Trizol, by Thermo Fisher Scientific (3 mentions)
Dynabeads protein g, by Thermo Fisher Scientific (3 mentions)
Rneasy minelute cleanup kit, by Qiagen (2 mentions)
Rip lysis buffer, by Solarbio (2 mentions)
Anti immunoglobulin g anti igg, by Merck Group (2 mentions)
Complete rip lysis buffer, by Merck Group (2 mentions)
Anti srsf1 antibodies, by Merck Group (2 mentions)
Control anti igg, by Merck Group (2 mentions)
91 protocols using anti ago2
1
RIP-based RNA Immunoprecipitation
2
Profiling Argonaute 2 Interactions in Lung Cancer
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After the A549 and NCI‐H1299 cell confluency reached 80%, the lung cancer cells were collected and lysed in the complete RIP lysis buffer. Cell extracts were subsequently incubated with magnetic beads conjugated with anti‐Argonaute 2 antibody (Anti‐Ago2; Millipore) or negative control normal mouse IgG (Millipore) overnight at 4°C, and were then digested with proteinase K (Invitrogen). The immunoprecipitated RNAs were purified with RNeasy MinElute Cleanup Kit (Qiagen), followed by analysis using RT‐qPCR assay. The assay was performed in triplicate.
3
RIP Assay for Circular RNA
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EZ-Magna RIP Kit (Millipore) was utilized for RIP assay. Briefly, A431 and SCL-1 cells were lysed with RIP buffer, and then the cell lysates were incubated with Protein A/G magnetic beads conjugated with anti-AGO2 (Millipore) or anti-IgG (Millipore). After washing with Proteinase K, the immunoprecipitated RNAs were isolated for detecting the enrichment of hsa_circ_0070934, miR-136-5p and PRAF2 using qRT-PCR.
4
RIP Assay for RNA-Binding Proteins
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The Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore) was used to conduct RIP assay. Briefly, HCCLM3 and MHCC97H cells were lysed in a RIP-lysis buffer. Then, the RIP buffer containing magnetic beads conjugated with anti-Ago-2 (Millipore) or anti-IgG (Millipore) were added to the cell lysates and incubated for overnight at 4°C. Next, the magnetic beads were incubated with proteinase K buffer for 30 min. Finally, the enrichment of HUMT, miR-455-5p, and LRP4 in purified RNAs was examined by qRT-PCR [41 ].
5
Immunoprecipitation of AGO2-Bound RNAs
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RNA immunoprecipitation (RIP) assays were executed by the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Burlington, MA, United States) according to manufacturer’s protocols. HRGC cells were lysed in lysis buffer and then incubated with RIP immunoprecipitation buffer which contained magnetic beads pre-incubated with the anti-AGO2 and anti-IgG (Millipore, United States). RNA was purified from RNA-protein complex and detected by qRT-PCR.
6
RIP Assay for miR-7-5p, RHPN1-AS1, and OGT
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EZ-Magna RIP kit (Millipore) was applied for conducting RIP assays. HCT-116 and HT29 cells were lysed in RIP lysis buffer, and incubated with anti-Ago2 (Millipore) and IgG (Millpore). The relative expression levels of miR-7-5p, RHPN1-AS1 and OGT were examined using qRT-PCR analysis.
7
Investigating miR-423-3p and CYBRD1 Interaction
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To determine the relationship between miR‐423‐3p and CYBRD1, Magna RIPTM RNA Binding Protein Immunoprecipitation Kit (Millipore) was introduced in RIP assay. In brief, RIP buffer was utilized for the lysis of NCI‐H1299 cells, and the cell extract was partly used as an input and partly incubated with anti‐Ago2 or anti‐IgG‐coated beads (Catalog No. 03–110; Millipore) 4°C for 6 h. After the cultivation of specimens with proteinase K buffer, immunoprecipitated RNA was isolated and then was purified. Finally, the CYBRD1 enrichment was measured by qRT‐PCR.
8
Ago2-RIP for RNA Profiling
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The Imprint RNA Immunoprecipitation Kit (Millipore, Bedford, OH, USA) was applied for RIP assay. Briefly, cell extracts were harvested after lysing in RIP lysis buffer (Solarbio, Beijing, China) and incubated with magnetic beads (Invitrogen) conjugated to antibodies of anti-Ago2 (Millipore) and anti-IgG (Millipore). RNA precipitates were extracted for qRT-PCR.
9
Ago2-Mediated RNA Immunoprecipitation
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RIP was implemented using a Magna RIP Kit (Millipore, Billerica, MA, USA) following the protocols. The beads were coated with human Ago2 antibody (anti-Ago2; Millipore) or mouse IgG antibody (anti-IgG; Millipore; control). Cells were lysed and cell lysates were incubated with anti-Ago2- or anti-IgG-coated beads. Afterwards, the coprecipitated RNAs were extracted and detected.
10
Quantifying RNA-Protein Interactions
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RNA immunoprecipitation (RIP) was analyzed using the Magna RIP RNA-binding protein immunoprecipitation kit (Millipore, Darmstadt, Germany) through Anti-AGO2 (#03-110, Germany). QRT-PCR was adopted to measure the RNA-bound complexes, and isotype control was conducted using anti-IgG.
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