A Magna RNA immunoprecipitation kit (Millipore) was adopted for RIP assay to confirm the associated relation between circ_0000463 and miR‐924. NSCLC cells were introduced with miR‐924 mimics for 48 h and then cells were disrupted using ice‐cold radio‐Immunoprecipitation Assay (RIPA) lysis buffer. Cell extracts were mixed with anti‐Argonaute 2 (Anti‐Ago2; Abcam) or anti‐Immunoglobulin G (Anti‐IgG; Abcam) for 4 h at 4℃. Then, the magnetic protein A/G beads were added to incubate for 2 h. Beads were washed with PBS thrice, and RT‐qPCR was applied to measure RNA enrichment.
Anti argonaute 2 anti ago2
Manufactured by Abcam
Sourced in United States
Anti‐Argonaute 2 (Anti‐Ago2) is a primary antibody that binds to the Argonaute 2 (Ago2) protein. Ago2 is a key component of the RNA-induced silencing complex (RISC) that plays a crucial role in the RNA interference (RNAi) pathway.
Automatically generated - may contain errors
Lab products found in correlation
Anti immunoglobulin g anti igg, by Abcam (5 mentions)
Magna rip rna binding protein immunoprecipitation kit, by Merck Group (3 mentions)
Proteinase k, by Solarbio (2 mentions)
Magna rna immunoprecipitation kit, by Merck Group (1 mentions)
Protease k, by Solarbio (1 mentions)
Magna rip kit, by Merck Group (1 mentions)
Rip lysis buffer, by Merck Group (1 mentions)
Anti ago2, by Abcam (1 mentions)
6 protocols using anti argonaute 2 anti ago2
1
RIP Assay for circ_0000463-miR-924 Interaction
2
Determination of miR-29c-3p and Foxo3 Enrichment
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Magna RNA-binding protein immunoprecipitation kits (EMD Millipore, Billerica, MA, USA) were applied for determination of RIP. In brief, KGN cells were lysed in RIP buffer and then cell extract (100 mL) was co-incubated with magnetic beads coated with Anti-Argonaute2 (Anti-Ago2; Abcam, Cambridge, MA, USA) or immunoglobulin G (Anti-IgG; Abcam) at 4°C for 8 h. The samples were then detached with protease K (Solarbio) for 30 min. Finally, RNA in the complex was isolated and then RT-qPCR was performed to assess the enrichment of miR-29 c-3p and Foxo3.
3
Confirming circRNF111-miR-27a-3p Interaction
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The Magna RIP RNA-binding Protein Immunoprecipitation Kit (Millipore, Bedford, MA, USA) was adopted to confirm the relationship between circRNF111 and miR-27a-3p. Cells were briefly lysed in RIP buffer and incubated with magnetic beads, which were conjugated with anti-Argonaute2 (anti-Ago2; Abcam) or anti-immunoglobulin G (anti-IgG; Abcam). Next, proteinase K (Solarbio) was added to digest the protein, and the RNA in the immunoprecipitated product was extracted. Finally, the co-precipitated circRNF111 and miR-27a-3p was detected by real-time qPCR after reverse transcription.
4
Confirming miR-199b-5p and circZNF124 Interaction
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The interaction between miR‐199b‐5p and circZNF124 or SLC7A5 was confirmed using a Magna RIP kit (Millipore). Briefly, the cultured HEC1A and Ishikawa cells were lysed with RIP lysis buffer (Millipore). The lysates were incubated with the magnetic beads coated with anti‐argonaute2 (anti‐Ago2; Abcam, Cambridge, UK) and anti‐immunoglobulin G (anti‐IgG; Abcam), respectively. After the beads were washed, both circZNF124 and miR‐199b‐5p enrichment in co‐precipitated RNAs were detected by qRT‐PCR.
5
Argonaute-2 Immunoprecipitation for circRNA
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RIP experiment was conducted utilizing the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Billerica, MA, USA). Briefly, A549 and H1299 cells were diluted in RIP buffer and maintained with Anti-Immunoglobulin G (Anti-IgG) or Anti-argonaute 2 (Anti-Ago2) (Abcam, Cambridge, MA, USA; Anti-Ago2 was used to detect Ago2, and then Ago2-RIP could be used to analyze the amount of circRNA and mRNA bound by miRNA) for 3 h. Then, the RNA in the immunoprecipitates was isolated and subjected to qRT-PCR as mentioned above.
6
RNA-Protein Interaction Profiling
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Magna RIP RNA Binding Protein Immunoprecipitation Kit (Millipore, Bedford, MA, USA) was adopted to confirm the relationship between miR‐140‐5p and circ‐RNF111 or E2F3. Briefly, PTX‐resistant cells were lysed in RIP buffer and incubated with magnetic beads which were conjugated with anti‐Argonaute2 (anti‐Ago2; Abcam, Cambridge, MA, USA) or anti‐immunoglobulin G (anti‐IgG; Abcam). Next, proteinase K (Solarbio) was added to digest the protein. Finally, immunoprecipitated RNA was isolated and the enrichment of circ‐RNF111, miR‐140‐5p and E2F3 was determined by qRT‐PCR.
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