RIP was conducted in SiHa cells with magna RIPTM RNA-binding Protein Immunoprecipitation kit (Millipore, Billerica, MA). SiHa cells were transfected with miR-320a mimics, and then lysed in complete RNA lysis buffer after 48 h. Cell lysates were rotated in RIP immunoprecipitation buffer including magnetic beads conjugated with negative control mouse IgG or human anti-AGO2 antibody (Mouse, Millipore, Billerica, USA) overnight. Next day, immunoprecipitated RNA was extracted after incubating with Proteinase K for 30 min. Last, qRT-PCR and agarose gel electrophoresis were performed to identify the expression of circCLK3 and miR-320a.
Human anti ago2 antibody mouse
Manufactured by Merck Group
Sourced in United States
The Human anti-AGO2 antibody (Mouse) is a laboratory reagent used for research purposes. It is a monoclonal antibody that specifically binds to the AGO2 protein, which is a component of the RNA-induced silencing complex (RISC) involved in gene silencing mechanisms. This antibody can be used to detect and study the expression and localization of AGO2 in various experimental systems.
Automatically generated - may contain errors
3 protocols using human anti ago2 antibody mouse
1
Investigating circCLK3 and miR-320a Interaction
2
Identifying circNHSL1 and miR-1306-3p interaction
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RIP assay was conducted with magna RIPTM RNA-binding Protein Immunoprecipitation kit (Millipore, Billerica, MA). MKN-28 cells were transfected with miR-1306-3p mimics or negative control. The cells were lysed in complete RNA lysis buffer after 48 h. Then, the RIP immunoprecipitation buffer including magnetic beads conjugated with negative control mouse IgG or human anti-AGO2 antibody (Mouse, Millipore, Billerica, USA) was added into cell lysates. Subsequently, the lysates were rotated overnight. Next day, after incubating with Proteinase K for 30 min, the immunoprecipitated RNA was extracted. Last, qRT-PCR and agarose gel electrophoresis were performed to identify the expression of circNHSL1 and miR-1306-3p.
3
Immunoprecipitation of RNA-Binding Proteins
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RIP assay was performed with Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Billerica, USA). In brief, 6×10 7 MGC-803 cells were lysed in 300μl complete RNA lysis buffer. Then, each 100μl cell lysates were added to 900μl RIP immunoprecipitation buffer containing magnetic beads conjugated with 5μg negative control mouse lgG, positive control anti-snRNP70 antibody or human anti-AGO2 antibody (Mouse, Millipore, Billerica, USA) and rotated overnight at 4℃. After incubation with proteinase K buffer at 55℃ for 30min, the immunoprecipitated RNA was puri ed and analyzed with RT-qPCR.
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