Electrophoretic Mobility Shift Assay (EMSA) Service

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Electrophoretic Mobility Shift Assay (EMSA) Service

Inquiry

With a mature electrophoretic mobility shift assay (EMSA) experimental platform and rich experience in nucleotide-protein-interacting analysis, CD BioSciences provides clients with EMSA service to qualitatively and quantitatively analyze whether plant proteins and DNA/RNA sequences interact.

Principle of EMSA

EMSA is a classical method for the research of plant transcription factors and cellular signal transduction pathways. It is based on the principle that DNA/RNA-protein complexes have different mobility in polyacrylamide gel electrophoresis (PAGE). Proteins bind to end-labeled nucleic acid probes, then PAGE, and the electrophoresis speed slows down after the protein binds to the probe. From the results of the PAGE gel, it is possible to determine, whether the protein binds to the nucleic acid sequence or not.

Figure 1. Schematic representation of RNA EMSA. – CD BioSciences Figure 1. Schematic representation of RNA EMSA. (Daras, G., et al., 2019)

Service Content

Workflow

  • Standard protein sample preparation and quantification.
  • Swimming lane setting and concentration gradient experimental design.
  • Probe design and synthesis.
  • Reaction of protein with probe.
  • EMSA gel electrophoresis migration assay.
  • Exposure imaging.
  • Experimental analysis and report writing.

Extension types of EMSA we provide

EMSA types Description
Validation EMSA This is used to verify that a probe contains a protein binding site.
Competitive EMSA To eliminate false positives and increase the accuracy of experimental results.
Super-shift EMSA Can verify the specific binding of the probe to the target protein.

Types of probes we can design

  • Isotope labeled probe

High specificity and sensitivity up to 10-18mol level.

  • Non-isotope labeled probes

Bioluminescent or chemiluminescent probes with high sensitivity and strong signals, without the radiography requirement.

Probe design

With many years of experience in probe design, we can

  • Prevent the probe from closing into a ring.
  • Prevent mispairing.
  • Exclude binding sites other than the target sequence.
  • Prevent spatial blocking.
  • Selection of appropriate AT/GC ratio.

Sample Requirements

Cells > 107 and plant tissues > 2g of protein to be extracted.

Cellular nuclear protein or purified transcription factor > 50uL/experimental group, concentration > 0.5 mg/mL, purity > 90%.

Probe sequence, if no probe sequence, please provide DNA binding protein-related information or related literature.

Binding protein-specific antibody > 50uL, concentration > 0.5mg/mL.

Target protein primary antibody and related kits.

Deliverables

  • Probe design and synthesis information.
  • Experimentally synthesized labeled probes.
  • Unlabelled probe (competitive EMSA).
  • Standard experimental flow report.
  • Original experimental pictures and data.

Please contact us for professional EMSA services. CD BioSciences, with its rich professional knowledge in plant biology and molecular technology, provides clients with the most advanced solutions for researching protein-nucleic acid interactions in plants. Collaborate with us, you will unravel the mechanisms of protein-nucleic acid interactions and facilitate the discovery of plant protein functions.

Reference

  1. Daras, G., et al. (2019). Detection of RNA-protein interactions using a highly sensitive non-radioactive electrophoretic mobility shift assay. Electrophoresis. 40(9), 1365-1371.

For research use only, not for clinical use.