Endogenous mir128-3p in the myocardium was detected with the miRCURY LNA miRNA Detection Probes (339111, Qiagen) on 6 µm paraffin sections. Sample preparation and hybridization were performed using miRCURY LNA miRNA ISH Buffer Set (339450, Qiagen). Briefly, the tissue was permeabilized with proteinase K for subsequent incubation with the double DIG-labelled LNA probe. Following stringent washing and blocking, samples were incubated with sheep anti-DIG AP FAB fragments (11093274910, Sigma). Freshly prepared AP substrate (11697471001, Roche) was applied for 2 hr at 30° C, which was followed by nuclear counterstaining with Nuclear Fast Red solution (N3020, Sigma). Consecutive sections were stained with H and E for morphological comparison. Brightfield images were captured using an Axiovision upright microscope equipped with a Coolsnap ES camera (Photometrics).
Mircury lna mirna ish buffer set
Manufactured by Qiagen
Sourced in Germany
The MiRCURY LNA miRNA ISH Buffer Set is a collection of reagents designed for the in situ hybridization (ISH) detection of microRNAs (miRNAs) in tissue samples. The set includes buffers and solutions necessary for the ISH workflow, providing the essential components for reliable and sensitive miRNA detection.
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2 protocols using mircury lna mirna ish buffer set
1
Detecting Endogenous miR-128-3p in Myocardium
2
FISH-based Cellular Localization
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Fluorescence-labeled probes (RiboBio) for RC3H2, 18S rRNA, and U6 RNA were designed and synthesized. The FISH experiments were performed using the Ribo fluorescent in situ hybridization kit (RiboBio). Images were obtained with a TCS SP2 laser-scanning confocal microscope (Leica Microsystems, Germany).
Cy3-labeled miR-101-3p probes (QIAGEN, Germany) was also synthesized. The cells were then prepared for immunofluorescence as described above. The FISH experiments were performed using miRCURY LNA miRNA ISH buffer set (QIAGEN, Germany). Images were obtained with a TCS SP2 laser-scanning confocal microscope (Leica Microsystems, Germany).
Cy3-labeled miR-101-3p probes (QIAGEN, Germany) was also synthesized. The cells were then prepared for immunofluorescence as described above. The FISH experiments were performed using miRCURY LNA miRNA ISH buffer set (QIAGEN, Germany). Images were obtained with a TCS SP2 laser-scanning confocal microscope (Leica Microsystems, Germany).
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