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.

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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

*Licensed from Roche Diagnostics GmbH

Figure 1

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

Table 1

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.

*Licensed from Roche Diagnostics GmbH

Figure 1