Analysis Biomolecular Interactions of P19 to E6/siRNA by BLI (CAT#: STEM-MB-0159-CJ)

Introduction

Protein-based systems for delivering siRNA can potentially circumvent some of the challenges facing nanoparticle-based systems, such as accumulation in the liver. Proteinaceous delivery vehicles commonly require both a carrier functionality, provided by a moiety that is chemically conjugated to or non-covalently complexed with siRNA, and an endosomal release functionality, which can be explicitly defined or embedded within the vehicle. p19 protein as an alternative siRNA-binding scaffold. Like dsRBDs, p19 binds specifically to double-stranded RNA (dsRNA) independent of sequence, and not to single-stranded RNA (ssRNA) or DNA. And p19 binds in a size-dependent manner to dsRNAs the length of siRNA, providing increased specificity. Importantly, p19 has a naturally higher affinity for siRNA, which provides an excellent backbone to further engineer ultra-high affinity siRNA carriers.

Add to Cart Please Inquire

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

Immunology/Inflammation; Pharmacology

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: Fortebio Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: FBS, DMEM (ATCC), Mouse Serum

Other recommended products

Analysis Biomolecular Interactions of NtAGO2 for KRAS by BLI (CAT#: STEM-MB-0195-CJ)
Analysis Kinetics of Intact Mab and scFv by BLI (CAT#: STEM-MB-0265-CJ)
Analysis Biomolecular Interactions of SARS-CoV-2 RBD and ACE2 by BLI (CAT#: STEM-MB-0101-CJ)
Analysis Biomolecular Interactions of PaParE and PaParD by BLI (CAT#: STEM-MB-0184-CJ)
Analysis Biomolecular Interactions of PTH-Fc and PTH1R by BLI (CAT#: STEM-MB-0152-CJ)
Analysis Biomolecular Interactions of IgG by Protein A by BLI (CAT#: STEM-MB-0131-CJ)
Analysis Biomolecular Interactions of TREM2 with LDL by BLI (CAT#: STEM-MB-0239-CJ)
Analysis Biomolecular Interactions of Cyclodextrin with Cholesterol by BLI (CAT#: STEM-MB-0140-CJ)