SOM Biophysics Facility
School of Medicine Biophysics Facility offer two biophysical analysis technologies;
- NanoSight NS300: It utilizes the particle characterization technology, Nanoparticle Tracking Analysis (NTA), to visualize and measure particles in suspension (aqueous solution only) in terms of size, light scattering intensity, fluorescence and count. NTA is a video-based analysis for particles between 20-2000 nm* (*material dependent). Its applications includes protein aggregation, exosome and microvesicle research, and drug delivery systems.
- MicroCal PEAQ (Isothermal Titration Calorimeter; ITC): This is the gold standard technique to study label-free biomolecular interactions in-solution and is a thermodynamic technique that directly measures the heat released or absorbed during a biomolecular binding event. It enables the accurate determination of binding constants (KD), reaction stoichiometry (n), enthalpy (ΔH) and entropy (ΔS). This provides a complete thermodynamic profile of the molecular interaction. The sample size for binder (e.g. proteins) is 300 ml and the MINIMUM requirement for the concentration of binder is 1 mM while that for the ligand is 10 mM. Both NanoSight and MicroCal PEAQ ITC are available to all School of Medicine, internal Deakin University researchers, as well as external scientists from other research facilities.
No training of these instruments will be provided.
Samples will be run by members of Prof. Wei Duan’s Lab.
- To gain access to these instruments, please provide a copy of SOP and complete the application form. This is to ensure that the experimental condition (eg. buffer used, nanoparticle waste management, etc.) is suitable and would not damage the instruments, and comply with regulatory requirements. Once the SOP is approved, booking can be arranged.
MicroCal PEAQ ITC
NanoSight NS300 (Nanoparticle Tracking Analysis) (KA4.308)
NanoSight NS300 utilize Nanoparticle Tracking Analysis (NTA) to characterize nanoparticles from 10nm - 2000 nm* (*dependent on material) in solution. Each particle is individually but simultaneously analysed by direct observations of diffusion. This particle-by-particle methodology produces high resolution results for particle size distribution and concentration while visual validation gives users extra confidence in the data. As well as particle size and concentration, protein aggregation and viscosity can all be analysed, while a fluorescence mode provides detection of labelled particles.
The NanoSight NS300 system uses a 405 nm (violet) laser source to excite suitable fluorophores whose fluorescence can then be determined using long-pass 430 nm emission filter.
Key Benefits of NanoSight NS300
MicroCal PEAQ (Isothermal Titration Calorimeter; ITC) (KA4.307)
Isothermal titration microcalorimeters measure the heat change that occurs when two molecules interact. Heat is released or absorbed as a result of the redistribution and formation of non-covalent bonds when the interacting molecules go from the free to the bound state. ITC monitors these heat changes by measuring the differential power, applied to the cell heaters, required to maintain zero temperature difference between the reference and sample cells as the binding partners are mixed. The reference cell usually contains water, while the sample cell contains one of the binding partners (the sample). A stirring syringe that holds the other binding partner (the ligand). The ligand is injected into the sample cell until the ligand concentration is two- to three-fold greater than the sample. Each injection of ligand results in a heat pulse that is integrated with respect to time and normalized for concentration to generate a titration curve of kcal/mol vs molar ratio (ligand/sample). The resulting isotherm is fitted to a binding model to generate the affinity (KD), stoichiometry (n) and enthalpy of interaction (ΔH). The MicroCal PEAQ-ITC is a highly sensitive, low volume isothermal titration calorimeter for the label-free, in-solution study of biomolecular interactions. It delivers direct measurement of all binding parameters in a single experiment and can analyze weak to high affinity binders, using as little as 10 µg sample.
Key Benefits of MicroCal PEAQ-ITC
MicroCal PEAQ-ITC analysis software
Booking and Cancellation
All bookings are to be made online using Outlook Calendar and are subjected to management approval. Minimum booking is 1 hour, followed by 30 minute increments thereafter.
*G HEALTH Medicine Lab Eqpmt ka 4.308 Nanosight300
*G HEALTH Medicine Lab Eqpmt ka 4.307 MicroCalPEAQITC
Charging starts from the time booked, so it is in your interest to arrive on time.
Cancellations can be made via the online booking system with between 24-48 hours’ notice.
Cancellations with less than 24 hours’ notice will be charged 50% of the booked time and no-shows will be charged in full.
- Malvern NanoSight NS300 Training Videos
- NanoSight NS300 Brochure
- Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis
- Exosomal microRNAs (exomiRs): Small molecules with a big role in cancer
- Particle size distribution of exosomes and microvesicles
- Measurement of Nanoparticles and Protein Aggregates
- Malvern ITC Training Videos
- MicroCal PEAQ-ITC: The gold standard for measuring binding affinities
- Principles of isothermal titration calorimetry (ITC)
- MicroCal PEAQ-ITC - Loading a Sample
- MicroCal PEAQ-ITC - How to clean the system
- MicroCal PEAQ-ITC - How to change the plunger tip
- MicroCal PEAQ-ITC - How to manually clean the syringe
- MicroCal PEAQ-ITC - How to replacing the syringe
- MicroCal PEAQ-ITC Brochure
- Analysis of Macromolecular Interactions by ITC
- ITC Review
- ITC Tutorial
- Introduction to Microcalorimetry Malvern
- ITC PEAQ DSC presentation April 2016_ATA
- Methods for measuring aptamer-protein equilibria: A review
- Methods for measuring small molecule-aptamer interaction ITC
- ITC Introduction Review 2010
- Characterization of biding interactions by ITC Review 1998
- Analysis of multiple-site binding of aptamer using ITC 2016
STANDARD OPERATING PROCEDURE
OCCUPATIONAL HEALTH & SAFETY
Tel. +61 3 5227 1149