Stem Cell-Associated miRNA Plate Array

Stem cells can differentiate into all of the specialized embryonic tissues during embryo development. In adult organisms, stem cells and progenitor cells act as a repair system for the body, replenishing specialized cells, in order to maintain the normal turnover of regenerative organs, such as blood, skin or intestinal tissues. Recent studies suggest that maintenance and differentiation of stem cells are regulated by microRNAs (miRNA). MiRNAs are small single-stranded RNA molecules (~20nt long), playing an important role in gene regulation and cell differentiation. MiRNAs are found to be differentially expressed in stem cells, such as brain-specific miR-124a and miR-9 molecules during neural lineage differentiation. In addition, miRNAs are also reported to regulate cancer stem cells. Signosis has developed a miRNA array specifically to target 47 miRNAs that are shown in literature to be associated with stem cell differentiation and maintenance. The array can be used for human, mouse and rat samples. Profiling the expression of these miRNAs will help to reveal the functions of miRNAs in stem cells.

A. Benefits:

Signosis' stem cell-associated miRNA plate array is a type of assays via direction hybridization of total RNA, which offers following benefits:

• Simple - Profiling the expression of 48 miRNA detection even simpler than ELISA. It does not need enzymatic conversion from miRNA into cDNA. The entire assay procedure is “incubation and wash”.

• Higher sensitivity and better discrimination power - The detection is approximately 100 times more sensitive than miRNA Northern blot. In addition, it has higher discrimination power than miRNA Northern blot in differetiating miRNA isoforms.

• Quantitative comparison – The difference of two or more samples in the expression of a specific miRNA can be quantitatively analyzed.

B. Principle of the technology

Signosis’ stem cell-associated miRNA plate array is a simple two-step assay; plate hybridization and streptavidin-HRP detection. The plate is pre-coated with an oligo mix, including a pair of unique oligos that hybridize side-by-side to a specific target miRNA and a universal capture oligo and a biotin-labeled oligo. In the assay, total RNA is directly utilized for hybridization. When the target miRNA exists in RNA, it acts as a bridge to bring the biotin-labeled oligo to the capture oligo, which can be detected through streptavidin-HRP conjugate and a chemiluminescent substrate. If there exists no specific miRNA, the biotin-labeled probe will be washed away, leading to no detection. In the plate array, 48 wells are coated with different oligo mixes for different miRNAs. A single 96-well plate allows quantitative measurement and comparison of 48 miRNAs between two samples. U6 RNA is used for normalization.

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