Analysis Biomolecular Interactions of DNA with β4-triad Mutant B3 Domains by BLI (CAT#: STEM-MB-0178-CJ)

Introduction

DNA with β4-triad mutant B3 Domains In plant and animal lineages that have independently evolved embryos , key gene regulatory networks that underlie complex development have evolved through functional diversification within families of related transcription factors. Functional diversification is facilitated by the typically modular domain architectures of transcriptional regulators. In plants, transcription factors that form a network controlling embryo formation and subsequent transition to the vegetative development share a conserved B3 DNA binding domain.In the B3 network of Arabidopsis, the AFL (ABI3, FUS3 and LEC2) B3 domain proteins primarily function as activators that promote embryogenesis.

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Principle Applications Procedure Materials Notes Related Products

Principle

Bio-Layer Interferometry (BLI) is an optical technique for measuring macromolecular interactions by analyzing interference patterns of white light reflected from the surface of a biosensor tip. BLI experiments are used to determine the kinetics and affinity of molecular interactions. In a BLI experiment, one molecule is immobilized to a Dip and Read Biosensor and binding to a second molecule is measured. A change in the number of molecules bound to the end of the biosensor tip causes a shift in the interference pattern that is measured in real-time.

Applications

Botany, Biochemistry

Procedure

1. Detect Buffers and prepare samples. BLI experiments are set up with one molecule immobilised on the surface of the biosensor (load sample) and a second molecule in solution (the analytical sample).
2. Fix the load sample on the biocompatible biosensor while the analytical sample is in solution.
3. The biosensor tip is immersed in the solution so that the target molecule begins to bind to the analysis sample.
4. Set up and run the BLI experiment. Molecules bound to or dissociated from the biosensor can generate response curves on the BLI system; unbound molecules, changes in the refractive index of the surrounding medium or changes in flow rate do not affect the interferogram pattern.
5. Collect and analyse data on the BLI's system.

Materials

• Equipment: Gator® Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: Directed Mutagenesis Kit

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