For setup and optimization of miRNA in situ hybridization (ISH) experiments

Features

Convenient setup and optimization of miRNA in situ hybridization (ISH) experiments

Superior sensitivity and specificity with double DIG-labeled detection probes and optimized reagents

Highly robust and supports high-throughput and individual miRNA localization studies

Fast and easy one-day protocol

High flexibility with no advanced instruments required

Validated in a wide range of tissues, including FFPE samples

Product Details

The miRCURY LNA miRNA ISH Optimization Kits (FFPE) offer all the essential reagents to set up and optimize miRNA in situ hybridization (ISH) experiments in the lab. These kits include double DIG-labeled miRCURY LNA miRNA Detection Probes, ISH buffer and reagents.

Performance

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).

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

Principle

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. 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.

Procedure

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.

Applications

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.