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miRCURY LNA™ microRNA ISH Optimization Kit (FFPE)

The miRCURY LNA™ microRNA ISH Optimization Kits (FFPE) offer all the essential reagents needed to set up and optimize microRNA in situ hybridization (ISH) experiments in the lab. They include double DIG*-labeled miRCURY LNA™ microRNA Detection Probes, ISH buffer and reagents. An easy-to-follow manual guides you through each of the steps of the experiment.
miRCURY LNA™ microRNA ISH Optimization Kit (FFPE)
  • The convenient way of setting up and optimizing microRNA in situ hybridization experiments in the lab
  • Superior sensitivity and specificity – Double DIG*-labeled miRCURY LNA™ microRNA Detection Probes and reagents specifically adapted to the kit
  • Very Robust – Advantageous for both high throughput and individual microRNA localization studies
  • Fast and easy – One-day microRNA ISH protocol
  • Highly flexible – No advanced instruments needed
  • Validated in a wide range of tissues – Ideal for use with clinical and experimental FFPE samples

Features

A miRCURY LNA™ microRNA ISH Optimization Kit (FFPE) is the ideal option for getting started with or optimizing microRNA in situ hybridizations (ISH) on formalin-fixed paraffin embedded (FFPE) tissue samples.

Based on the highly successful double (5’ and 3’) DIG-labeled miRCURY LNA™ microRNA Detection Probes, the kits provide the sensitivity and specificity needed to detect microRNAs. In addition, the kits ship with reagents specifically adapted for use with LNA™ probes in FFPE tissue sections.

The accompanying instruction manual carefully explains each step of the ISH experiment and provides tips and recommendations for a successful experiment. Furthermore, it includes a thoroughly validated one-day protocol for fast and trouble-free ISH analysis.

Applications

The kits can be used for a large number of applications including cellular and sub-cellular microRNA localization studies and determination of spatial microRNA expression (for more applications, click on the Experimental data tab). Furthermore, the robustness of the procedure makes it ideal for use in both high-throughput ISH analysis and localization studies of individual microRNAs.

Coverage

Eight different miRCURY LNA™ microRNA ISH Optimization Kits are available. Each kit comes with positive and negative control probes, hybridization buffer and Proteinase K. In addition, a unique tissue-specific miRCURY LNA™ microRNA Detection Probe is included in each kit.

Each kit includes:
  • Unique microRNA LNA™ probe (double DIG-labeled, kit specific)
  • Scrambled LNA™ probe (double DIG-labeled, negative control)
  • U6 LNA™ probe (5’ DIG-labeled, positive control)
  • Hybridization buffer (2x, formamide-free)
  • Proteinase K
The unique microRNA LNA™ probes have been validated in a variety of tissues and are therefore ideal for optimizing ISH experimental settings. Use Table 1 for recommendations on which kit to choose based on tissue type.

Table 2 lists the unique probes in each kit and provides examples of where the targeted microRNA is expressed.

Table 2. Description of the kits.
Product Unique probe Examples of miR-expressing cells
Image
Kit 1 miR-1 Skeletal muscle cells, myocytes, myocardiocytes
View image
Kit 2 miR-21 Cancer-associated fibroblasts, some immune cells
View image
Kit 3 miR-122 Hepatocytes
View image
Kit 4 miR-124 Neurons
View image
Kit 5 miR-126 Endothelial cells
View image
Kit 7 miR-145 Smooth muscle, vascular smooth muscle (pericytes), myoepithelial cells
View image
Kit 8 miR-205 Basal keratinocytes, myoepithelium, SCC cancer cells
View image
Kit 9 miR-223 Myeloid, granulocytic, and monocytic compartments of the hematopoietic system
View image


 
Figure 1 Principle of the miRCURY LNA™ microRNA ISH Optimization Kit (FFPE)
Principle of the miRCURY LNA™ microRNA ISH Optimization Kit (FFPE). (Click to learn more)


Table 1 Kit recommendations based on tissue types
Kit recommendations based on tissue types. (Click to learn more)

Double DIG*-labeled probes for superior sensitivity

The miRCURY LNA™ microRNA ISH Optimization Kits (FFPE) use double DIG-labeled miRCURY LNA™ microRNA Detection Probes. These probes offer exceptional sensitivity and specificity which means that they even can be used for detection of less abundant microRNAs.

All the kits have been thoroughly validated in a wide range of tissues to ensure that they offer the highest level of performance (Figures 1-7).

The ideal way of optimizing microRNA ISH experiments

All kits ship with a double DIG-labeled scrambled probe and a 5’ DIG-labeled U6 control probe. The U6 probe targets U6 snRNA, a small RNA expressed in most tissues, which makes it the ideal positive control probe in the initial stages of setting up an ISH experiment. The scrambled probe is used as a negative control probe.

In addition to these probes, a kit-specific double DIG-labeled miRCURY LNA™ microRNA Detection Probe is provided with each kits. These probes can be used as positive control probes in the experiment, once the initial optimization has been performed. Taken together, the kits provide everything you need for getting started with FFPE microRNA ISH experiments.

A solution for many problems

The miRCURY LNA™ microRNA ISH Optimization Kits (FFPE) are very flexible, offering solutions for many different experimental setups without the need for advanced instruments. Some of the applications are listed below.

  Kit applications
  • The study of cellular and sub-cellular microRNA localization.
  • Examination of spatial microRNA expression.
  • MicroRNA expression analysis during developmental processes.
  • ISH analysis of clinical FFPE samples.
  • ISH analysis of experimental FFPE samples.
  • Diagnostic and prognostic biomarker discovery in clinical FFPE samples.
  • Potential microRNA ISH in other species where microRNA target sequence for kit-specific LNA™ probe is conserved.
  • Evaluation of results from functional studies.

A robust procedure

The microRNA ISH protocol that comes with the kit has been thoroughly tested and optimized. Exiqon’s scientists have managed to eliminate several of the steps normally associated with ISH, such as pre-hybridization, post-fixation and acetylation, thus making the protocol very robust, easy to optimize and ideal for high throughput use.

Non-toxic reagents

Both the hybridization and washing steps of Exiqon’s ISH protocol are performed using completely formamide-free buffers, which makes working with the kits less cumbersome.



Figure 1 miR-1 detection (kit 1) in adult mouse heart
miR-1 detection (kit 1) in adult mouse heart. (Click to learn more)

Figure 2 miR-21 detection (kit 2) in colon adenocarcinoma
miR-21 detection (kit 2) in colon adenocarcinoma. (Click to learn more)

Figure 3 miR-122 detection (kit 3) in mouse liver
miR-122 detection (kit 3) in mouse liver. (Click to learn more)

Figure 4 miR-124 detection (kit 4) in human brain
miR-124 detection (kit 4) in human brain. (Click to learn more)

Figure 5 miR-126 detection (kit 5) in breast cancer
miR-126 detection (kit 5) in colon wall. (Click to learn more)

Figure 6 miR-145 detection (kit 7) in human colon
miR-145 detection (kit 7) in human colon. (Click to learn more)

Figure 7 miR-205 detection (kit 8) in human breast carcinoma
miR-205 detection (kit 8) in human breast carcinoma. (Click to learn more)

Figure 8 miR-223 detection (kit 9) in esophagus cancer
miR-223 detection (kit 9) in esophagus cancer. (Click to learn more)
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