MIR31 in situ Hybridization Assay

The MIR31 in situ hybridization assay was performed as described previously with modifications (Tian Y. et al., 2017 (link)). Digoxigenin-labeled LNA probes (Exiqon, Vedbaek, Denmark) were used following the manufacturer’s protocol. Both digoxigenin-labeled MIR31 and scrambled LNA probes (Exiqon) were hybridized at 61°C. The U6 probe was used as a positive control. In situ signals were detected by staining with anti-digoxigenin-AP antibody (Roche, Basel, Switzerland) and developed using BM purple substrate (Roche).

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Qu J., Shao C., Ying Y., Wu Y., Liu W., Tian Y., Yin Z., Li X., Yu Z, & Shuai J. (2022). The spring-like effect of microRNA-31 in balancing inflammatory and regenerative responses in colitis. Frontiers in Microbiology, 13, 1089729.

Publication 2022

Digoxigenin-labeled LNA probes scrambled LNA probes anti-digoxigenin-AP antibody BM purple substrate

Anti antibody Assay Digoxigenin In situ hybridization

Corresponding Organization : China Agricultural University

Other organizations : Fujian Jiangxia University, Zhejiang Lab, University of Chinese Academy of Sciences, Wenzhou University

Top 5 similar protocols

Variable analysis

independent variables

Hybridization temperature

dependent variables

MIR31 expression (in situ signals)

control variables

Digoxigenin-labeled LNA probes
Digoxigenin-labeled scrambled LNA probes
Anti-digoxigenin-AP antibody
BM purple substrate

positive control

U6 probe

negative control

Digoxigenin-labeled scrambled LNA probes

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