RIP Assay Verifies miR-214-5p Binding

RIP was used to verify the binding relationship between miR-214-5p and PSMA3-AS1 or PD-L1 [22 (link)]. RIP assay was conducted with an EZMagna RIP kit (Merck KGaA). Cell lysates were treated with magnetic beads conjugated to Ago2 antibody (Abcam) or control IgG antibody (Abcam) in RIP buffer. All precipitated RNAs were examined by RT-qPCR.

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Zhang M., Xu Y., Yin S, & Qiu F. (2021). YY1-induced long non-coding RNA PSMA3 antisense RNA 1 functions as a competing endogenous RNA for microRNA 214-5p to expedite the viability and restrict the apoptosis of bladder cancer cells via regulating programmed cell death-ligand 1. Bioengineered, 12(2), 9150-9161.

Publication 2021

EZMagna RIP kit Ago2 antibody control IgG antibody

Ago2 Antibody Assay Buffer Cell Igg antibody Pd l1 Rnas

Corresponding Organization : First Affiliated Hospital of Soochow University

Other organizations : The First People's Hospital of Changzhou, First People's Hospital of Kunshan, Jiangsu University, Xian Yang Central Hospital

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Variable analysis

independent variables

Treatment of cell lysates with magnetic beads conjugated to Ago2 antibody
Treatment of cell lysates with magnetic beads conjugated to control IgG antibody

dependent variables

Expression levels of RNAs precipitated during the RIP assay

control variables

RIP buffer

controls

Positive control: Ago2 antibody
Negative control: Control IgG antibody

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