At BMCU, we are equipped with a number of instruments to probe biomolecular interactions.
These include optical (surface plasmon resonance) and gravimetric (quartz crystal microbalance with dissipation) biosensing instruments, as well as fully automated isothermal calorimetry. In addition, using our LigandTracer® Green instrument we can do real-time quantification of binding of fluorescently labeled biomolecules or bacterial cells to immobilized cells or receptors. We can assist with measurement of interactions of proteins with other proteins, RNA, lipids carbohydrates or small molecules.
For further information you can contact Johan Olofsson Edlund, or the person responsible for the instrument you are interested in (listed below together with the instrument).
Biacore3000
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Location The room is: Building KBC, 6th floor, room A5 39 07. Directions using digital map
About Surface plasmon resonance (SPR) is an optical biosensing method which probes the adsorption of biomolecules onto surfaces by monitoring, in real time, changes in the refractive index occurring at the sensing interface. This change in refractive index can be related to the mass adsorbed onto the chip. In an SPR-based measurement to probe biomolecular interactions, one of the targets (the ligand) is attached onto the chip while the other ligand (target) is flown over the chip. A number of chips with different chemical modifications are commercially available and allow ligand immobilization e.g. through his-tags, biotin-tags or covalent coupling via amines.
SPR has become one of the most widely adopted surface-based techniques for measuring the affinity of biomolecules. It can be used, for example to probe protein-small molecule, protein-carbohydrate, protein-lipid membrane or protein -RNA interactions. With the kinetic assays you can determine association rate constant, dissociation rated constants and affinity the interaction.
The Biacore 3000 is an older instrument which gives the user the option of doing “manual” injections to better optimize running conditions and also to create own scripts for reproducible workflows. Less experienced users can also use the application wizard templates and be guided through experiments step by step with the internal software.
Proteon XPR36
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Location The room is: Building KBC, 6th floor, room A5 39 07 Directions using digital map
About The ProteOn XPR36 is an SPR based biosensor from BIO-RAD. The measuring principles are the same as for the Biacore3000 system in the regards that it is measuring the change in mass bound to the chip. The main difference to the Biacore system is that it is capable of 6 simultaneous injection over 6 different flowcell surfaces. The other key difference is the thickness of the capillaries in the systems, the ProteOn has wider capillaries and is therefore more robust and less sensitive to clogging. The trade off for this is that the sensitivity is slightly lower in comparison to the Biacore.
Quartz Crystal Microbalance with Dissipation monitoring (QCM-D)
Quartz Crystal Microbalance with Dissipation monitoring (QCM-D)
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Location The room is: Building 6M, 4th floor, room 6070 6070, 6M, Norrlands universitetssjukhus Nus, digital map
Price 500 SEK/day Introduction/driver license Cost is 1500 SEK
About Quartz Crystal Microbalance with Dissipation monitoring (QCM-D) is a label free technique that allows for the real-time monitoring of changes in both mass adsorbed to the sensor surface and in the viscoelastic properties of materials adsorbed to the sensor surface. While this instrument can be used for a wide variety of experiments ranging from materials science to health care, the host lab commonly uses it to study how proteins interact with lipid membranes and/or for investigating protein-protein interactions. While SPR instruments (also available at BICU) provide the dry mass of the adsorbed protein, this instrument measures the hydrated mass which when combined with the dissipation monitoring allows, among other, changes in packing density of proteins interacting with the surface to be resolved.
Price 1000 SEK/day Introduction/driver license Cost is 1000 SEK (half a day)
About Auto-ITC200 is a fully automatec isothermal caliometry system that measures affinity based on heat exchange when two molecules interact. It can be used to determine the dissociation constant, enthalpy, entropy and stoichiometry of interactions. The most common applications are protein-protein and protein-small molecule interactions, but it will be able to measure any type of interaction that gives of a large enough signal. IN comparison with the SPR systems the ITC measures in solution and not bound on surfaces, so while you may need more materal compared to the SPR approach, you will not need to worry about any surface immobilizaton.
Price 1000 SEK/day Introduction/driver license Cost is 1000 SEK (half a day)
About ITC200 is a manual isothermal caliometry system that measures affinity based on heat exchange when two molecules interact. As with the Auto-ITC200, it can be used to determine the dissociation constant, enthalpy, entropy and stoichiometry of interactions.
The most common applications are protein-protein and protein-small molecule interactions, but it will be able to measure any type of interaction that gives of a large enough signal. In comparison with the SPR systems the ITC measures in solution and not bound on surfaces, so while you may need more materal compared to the SPR approach, you will not need to worry about any surface immobilizaton.
The advantage of the manual ITC200 vs the Auto-ITC200 is that it consumes slightly less sample than the automated one. The downside is that you need to perform each run manually.
Price 500 SEK/day Introduction/driver license Cost is 1500 SEK
About LigandTracer® Green – an instrument for real-time quantification of binding of fluorescently labeled biomolecules or bacterial cells to immobilized cells or receptors.
LigandTracer® Green offers a simple and accurate method for detection and quantification of biomolecular/cells interactions in real‐time using fluorescently labeled biomolecules. Measurements of e.g. on‐off rate, affinity or retention can be made using blue (~488nm) and green (~535nm) fluorophores, e.g. FITC, Alexa Fluor® 488, as labels. A detector for Yellow (590nm) - Red (632nm) for Rhodamine Red and similar fluorophores is also available.