RIP assay was conducted as described by Chen et al.22 (link) The Imprint® RNA immunoprecipitation kit (Sigma) was used for RIP assay in compliance with recommendations. Antibodies used in the RIP assay were bought from Millipore (Billerica, MA, USA), including Argonaute-2 (Ago2; Millipore) or IgG (Millipore) antibodies. Furthermore, HPMECs were transfected with bio-miR-31-5p-wt, bio-miR-31-5p-mut, or bio-miR-NC (RiboBio) at a final concentration of 50 nM for 48 h before harvest. Cell lysates were treated with streptavidin-coated magnetic beads overnight at 4 °C. After centrifugation at 10,000×g for 10 min, RNA complexes bound to the beads were extracted for RT-qPCR assay.
Argonaute2 ago2
Manufactured by Merck Group
Sourced in United States
Argonaute2 (Ago2) is a laboratory equipment product offered by the Merck Group. Ago2 is a protein that is a key component of the RNA-induced silencing complex (RISC), which plays a central role in the RNA interference (RNAi) pathway. Ago2 binds to small interfering RNAs (siRNAs) or microRNAs (miRNAs) and uses them to identify and cleave target mRNA molecules, thereby regulating gene expression.
Automatically generated - may contain errors
Lab products found in correlation
Bio mir nc, by RiboBio (1 mentions)
Imprint rna immunoprecipitation kit, by Merck Group (1 mentions)
Ez 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)
Rnase inhibitor, by Merck Group (1 mentions)
Immunoglobulin g igg, by Merck Group (1 mentions)
3 protocols using argonaute2 ago2
1
RIP Assay for miRNA-mRNA Interactions
2
Argonaute2-Mediated circVMP1 Regulation
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A commercial EZ-Magna RIPTM RNA-Binding Protein Immunoprecipitation Kit (Millipore) was used to perform RIP assay. Cell extracts were mixed with beads coated with Argonaute2 (Ago2; Millipore) or Immunoglobulin G (IgG; Millipore) antibody. The enrichment of circVMP1, miR-524-5p, METTL3, and SOX2 was analyzed by RT-qPCR. RIP assay was performed three times.
3
Investigating miR-1253 and hsa_circ_0000190 Interaction
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To confirm the ability of miR-1253 and hsa_circ_0000190 to interact directly with one another, the EZ-Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Millipore) kit was used based on provided instruction. Lysis buffer supplemented with RNase inhibitor (Millipore) was used for the initial preparation of cell lysates, which were subsequently incubated with Argonaute2 (Ago2; Millipore) or Immunoglobulin G (IgG; Millipore) antibody-coated magnetic beads. A qPCR approach was subsequently used to quantify RNA enrichment.
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