The detection of miR135b-5p expression in tissue sections was performed using the miRCURY LNA microRNA Detection Kit (QIAGEN, Hilden, Germany) as described.32 (link) Staining was performed on formalin-fixed, paraffin-embedded patient tissues. Briefly, the sections were dewaxed in Roti-Histol (Carl Roth, Karlsruhe, Germany, rehydrated in 2-propanol, treated with proteinase K (15 mg/mL; QIAGEN, Hilden, Germany) and air dried. Hybridization was performed for 2 h at 52°C, using a miR-135b-5p-specific sequence; the sequence of the digoxigenin-labeled locked nucleic acid (LNA) detection probe was 5′- TCACATAGGAATGAAAAGCCA-3′. A scrambled miRNA probe was used as a negative control. After stringent washes, the bound LNA probes were detected with alkaline phosphatase-conjugated digoxigenin Ab (Roche Diagnostics, Mannheim, Germany) and nitroblue tetrazolium/5-bromo-4-chloro-3'-indolyphosphate p-toluidine (NBT/BCIP; Vector Laboratories, Burlingame, CA, USA) as the substrate. Nuclear Fast Red (Vector Laboratories, Burlingame, CA, USA) was used for nuclear staining. The sections were mounted using Roti-Mount FluorCare.
Mircury lna microrna detection kit
Manufactured by Qiagen
Sourced in Germany
The MiRCURY LNA™ microRNA Detection Kit is a laboratory product designed for the detection and quantification of microRNA (miRNA) expression. It utilizes Locked Nucleic Acid (LNA) technology to provide sensitive and specific detection of miRNA targets.
Automatically generated - may contain errors
Lab products found in correlation
Fast red, by Vector Laboratories (2 mentions)
Roti histol, by Carl Roth (2 mentions)
Nuclear fast red, by Vector Laboratories (1 mentions)
Nbt bcip, by Vector Laboratories (1 mentions)
Proteinase k, by Qiagen (1 mentions)
Rnaimax, by Thermo Fisher Scientific (1 mentions)
Nbt bcip, by Thermo Fisher Scientific (1 mentions)
Roti mount fluorcare, by Carl Roth (1 mentions)
5 protocols using mircury lna microrna detection kit
1
Detection of miR135b-5p in Tissue Sections
2
H19 lncRNA Detection in PDAC Tissues
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The miRCURY LNA™ microRNA Detection Kit (QIAGEN, Hilden, Germany) was used according to the instructions of the manufacturer. A H19 probe, which was labeled with digoxigenin (DIG) at the 3′ and 5′ ends, was hybridized to PDAC tissue at 54 °C for 2 h. The sequence of the H19 probe was 5′-AATGCTTGAAGGCTGCTCCGT-3′. After stringent washes, the bound H19 probe was detected with nitroblue tetrazolium/5-bromo-4-chloro-3′-indolyphosphate p-toluidine (NBT/BCIP; Vector Laboratories, Burlingame, CA, USA), which served as a substrate. For nuclear staining, the slices of patient tissue were incubated in Fast Red (Vector Laboratories, Burlingame, CA, USA) for 1 min.
3
miRNA Expression Analysis in PDAC
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The miRCURY LNA™ microRNA Detection Kit (QIAGEN, Hilden, Germany) was used to detect the expression of miRs in PDAC tissue sections according to the instructions of the manufacturer. The hybridization was performed for 2 h at 54 °C using a specific 3′,5′ digoxigenin (DIG)‑labeled LNA miR detection probe. A scrambled, DIG-labeled miR served as a negative control. The sequences of the miR probes were as follows: hsa-miR-378i: 5′-DiGN-CCTTCTGACTCCTAGTCCAGT-3′-DiGN_N; hsa-miR-378a-3p: 5′-DiGN-CCTTCTGACTCCAAGTCCAGT-3′-DiGN_N. The bound miR probes were detected with nitroblue tetrazolium/5-bromo-4-chloro-30-indolyl phosphate p-toluidine (NBT/BCIP; Vector Laboratories, Burlingame, CA, USA), which served as a substrate. For nuclear staining, patient tissue slices were incubated in Fast Red (Vector Laboratories, Burlingame, CA, USA). Positive signals in ten randomly selected fields were counted blindly by two examiners.
4
Quantifying miR-21 Expression in Gastric Cancer
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qRT-PCR was performed for miR-21 as described above. CISH was performed on FFPE tissue sections using a miRCURY LNA microRNA detection kit (Exiqon, details in supplementary materials). For the miR-21 transfection study, 15-PGDH-directed miR-21 (UAGUCAGACUAUUCGAu) was constructed as previously described17 (link). The miR-21 and scramble mimic were transfected into gastric cancer cells using RNAiMAX (Invitrogen) according to the manufacturer’s instructions.
5
miR-132 Expression Detection in Tissues
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Detection of miR-132 expression in tissue sections was achieved using miRCURY LNA™ microRNA Detection Kit (Exiqon, Vedbaek, Denmark) as described (32 (link)). Staining was performed on formalin-fixed, paraffin-embedded patient tissues. Briefly, sections were dewaxed in Roti-Histol (Carl Roth GmbH & Co. KG, Karlsruhe, Germany), rehydrated in 2-propanol, treated with proteinase K (15 µg/ml) and air-dried. Hybridization was performed for 2 h at 58°C using a miR-132-specific, digoxigenin-labeled locked nucleic acid (LNA) detection probe and a scrambled miR as a negative control. After stringent washes, the bound LNA probes were detected with an alkaline phosphatase-coupled digoxigenin antibody (Sigma-Aldrich; Merck KGaA) and NBT/BCIP (Thermo Fisher Scientific, Inc.) as the substrate. The sections were mounted using Roti-Mount FluorCare (Carl Roth GmbH& Co. KG) containing 4′,6-diamidino-2-phenylin-dole (DAPI) as a counterstain.
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