RNA Immunoprecipitation (RIP) Assay
This protocol is extracted from research article:
Novel lncRNA LINC00941 Promotes Proliferation and Invasion of Colon Cancer Through Activation of MYC
Onco Targets Ther, Feb 22, 2021; DOI: 10.2147/OTT.S293519

RIP assay was performed using Magna rip RNA binding protein immunoprecipitation Kit (Millipore, Billerica, Ma, USA). Simply, the transfected colon cancer cells were collected, and lysed in lysis buffer. Then the cell lysate and RIP buffer containing magnetic-AgO2 complex (the magnetic beads were pre-incubated with AgO2 antibody at room temperature) were incubated at 4 °C. Magnetic beads were coupled with anti-IgG as a negative control. Thereafter, immunoprecipitated RNA was extracted and the AgO2-bound RNA expression was measured by qRT-PCR.

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