RNA Pulldown

Technologys

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RNA Pulldown

A growing number of researches have shown that non-coding RNAs have powerful and diverse biological effects. However, little is known about the mechanisms of non-coding RNA function. To understand these mechanisms, it is important to define the interactions between ncRNA and cellular proteins. The identification of these ribonucleoprotein complexes is a key step in understanding how non-coding RNAs function. In vitro RNA pulldown techniques can be used to identify protein chaperone interests that interact with RNA.

The RNA pulldown assay selectively extracts protein-RNA complexes from the sample. In general, RNA pulldown assays take advantage of high-affinity tags (such as biotin, etc.) The RNA probe can be biotinylated to complex with proteins in the cell lysate and then purified using agarose or magnetic beads. Alternatively, the protein can be labeled or the RNA-protein complex can be isolated using an antibody to the protein of interest. RNA is then detected by Northern blot or RT-PCR analysis, and protein is detected by Western blot or mass spectrometry.

IntegrateRNA provides you with optimized RNA pulldown services, including biotin labeling and antibody labeling, to suit your interaction research project.

Strategy of our RNA pulldown

  • Determine the research plan according to the purpose of the research
  • Construction and validation of labeled RNA expression vectors (construct captured protein expression vectors for antibody labeling)
  • Production of labeled RNA probes by in vitro transcription
  • Elution and separation of pulldown products (including ncRNA, mRNA, RBP)
  • Pull down product analysis (Western blot and mass spectrometry)
  • Data analysis and graphics production

Final delivery

  • Gene synthesis sequencing report
  • Construction of a plasmid containing the gene of interest, bacterial solution, sequencing report
  • In vitro transcription template for sense & antisense primers, template electropherogram
  • RNA pulldown protein SDS-PAGE silver staining
  • Mass spectrometry report

Applications

  • Study on RNA functional mechanism
    ✧ Mutation analysis
    ✧ Structural function analysis
    ✧ Identification of RNA: protein interactions
    ✧ MS analysis of unknown RNA — protein binding pairs or complexes
  • High-throughput screening of potential interacting molecules

IntegrateRNA offers a variety of in vivo and in vitro systems for RNA functional studies to help our customers optimize project research solutions to increase success rates and reduce cycle times. We guarantee that all services and products are subject to strict quality control. Based on professional, efficient and experienced services, IntegrateRNA has won the unanimous trust and support of customers. If you have any questions, please feel free to contact us.

For research use only. Not intended for any clinical use.
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