miRCURY LNA miRNA ISH Optimization Kits (FFPE)

Double DIG-labeled probes for superior sensitivity

The miRCURY LNA miRNA ISH Optimization Kits (FFPE) use double DIG-labeled miRCURY LNA miRNA Detection Probes. These probes offer exceptional sensitivity and specificity, so they even can be used to detect low-abundance miRNAs.

All of the kits have been thoroughly validated for use with a wide range of tissues to ensure the highest level of performance (see table below).

Description of the miRCURY LNA miRNA ISH Optimization Kits (FFPE)

Product	Unique probe	Examples of miR-expressing cells	Data figure
Kit 1	miR-1	Skeletal muscle cells, myocytes, myocardiocytes	miR-1 detection in adult mouse heart
Kit 2	miR-21	Cancer-associated fibroblasts, some immune cells	miR-21 detection in colon adenocarcinoma
Kit 3	miR-122	Hepatocytes	miR-122 detection in mouse liver
Kit 4	miR-124	Neurons	miR-124 detection in human brain
Kit 5	miR-126	Endothelial cells	miR-126 detection in colon wall
Kit 7	miR-145	Smooth muscle, vascular smooth muscle (pericytes), myoepithelial cells	miR-145 detection in human colon
Kit 8	miR-205	Basal keratinocytes, myoepithelium, SCC cancer cells	miR-205 detection in human breast carcinoma
Kit 9	miR-223	Myeloid, granulocytic, and monocytic compartments of the hematopoietic system	miR-223 detection in esophagus cancer

The ideal way to optimize miRNA ISH experiments

All of the miRCURY LNA miRNA ISH Optimization Kits (FFPE) include 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, making it an 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, a kit-specific, double DIG-labeled miRCURY LNA miRNA Detection Probe is provided with each kit. These probes can be used as positive control probes in the experiment, once the initial optimization has been performed.

A solution for many problems

The miRCURY LNA miRNA ISH Optimization Kits (FFPE) are very flexible, offering solutions for many different experimental setups without the need for advanced instruments. Some applications include:

• Study of cellular and sub-cellular miRNA localization
• Examination of spatial miRNA expression
• miRNA expression analysis during developmental processes
• ISH analysis of FFPE samples
• Biomarker discovery in FFPE samples
• Potential miRNA ISH in other species where the miRNA target sequence for the kit-specific LNA probe is conserved
• Evaluation of results from functional studies

Features

The miRCURY LNA miRNA ISH Optimization Kit (FFPE) is a great tool to help you get started with or to optimize miRNA 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 miRNA Detection Probes, the kits provide the sensitivity and specificity needed to detect miRNAs (see figure Principle of the miRCURY LNA miRNA ISH Optimization Kit). In addition, the reagents included have been specifically adapted for use with LNA-enhanced probes in FFPE tissue sections.

Coverage

Eight different miRCURY LNA miRNA 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 miRNA Detection Probe is included in each kit.

Each kit includes:

• Unique miRNA 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 miRNA LNA probes have been validated in a variety of tissues and are therefore ideal for optimizing ISH experimental settings. For recommendations on which kit to choose based on tissue type, refer to the figure miRCURY LNA miRNA ISH Kit chooser.

The One-Day miRNA ISH Protocol included in the kit handbook has been thoroughly tested and optimized. Our scientists have eliminated several steps normally associated with ISH, such as pre-hybridization, post-fixation and acetylation, making the protocol very robust, easy to optimize and ideal for high-throughput use. Both the hybridization and washing steps of the One-Day miRNA ISH Protocol are performed using completely formamide-free buffers, which makes working with the kits less cumbersome.

The miRCURY LNA miRNA ISH Optimization Kits (FFPE) can be used for a large number of applications, including cellular and sub-cellular miRNA localization studies and determination of spatial miRNA expression. The robustness of the procedure makes it ideal for use in both high-throughput ISH analysis and localization studies of individual miRNAs.