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RNA Electrophoretic Mobility Shift Assay


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RNA Electrophoretic Mobility Shift Assay Inquiry

As a leader specialized in RNA-protein interaction analysis, Lifeasible has acquired extensive experience and expert knowledge in RNA-protein interaction study. RNA electrophoretic mobility shift assay (EMSA) is an in vitro technique commonly employed in a primary characterization of RNA-protein interactions. Lifeasible offers RNA EMSA service with high sensitivity and efficiency.

The RNA EMSA, also known as the gel retardation assay, takes advantage of the fact that protein-associated nucleic acids migrate more slowly than free probes in native polyacrylamide gel (Figure 1).

Gel_shift_assay.png
Figure 1. The principle of an RNA EMSA (Wikipedia)

Lane 1 is a negative control and contains only genetic material. Lane 2 contains protein as well as an RNA fragment that does not interact. Lane 3 contains protein and an RNA fragment that does interact, causing a shifted band.

Generally, a conventional RNA EMSA protocol consists of five steps, including:

  • Preparation of a protein sample. A protein sample is typically from a cell lysate or expression in vitro. We provide optimized solutions for protein extraction or expression.
  • Synthesis and labeling of a nucleic acid. Lifeasible provides RNA probes labeled with radioisotopes, fluorescent, and chemiluminescent to meet the diverse requirements of customers.
  • A binding reaction. A labeled RNA probe is incubated with a protein sample to initiate binding and formation of an RNA-protein complex.
  • Native gel electrophoresis. The RNA-protein complex is separated via non-denaturing polyacrylamide gel electrophoresis.
  • Detection. An RNA-protein complex migrates slower than a free RNA probe through a gel matrix, causing a migration shift relative to the unbound RNA probe. The shifted band can be detected by autoradiography, fluorescence, or chemiluminescence methods.

We provide one-site RNA EMSA service from sample preparation to data interpretation. We will be happy to discuss details of intended RNA-protein interaction studies with you and develop experimental strategies tailored to your requirement. Please feel free to contact us for any further information on our services or assistance you might need.

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