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Affinity Measurements

We can assist with measurement of interactions of proteins with other proteins, RNA, lipids and carbohydrates. We can also perform fully automated iso thermal calorimetry. In addition, we can do real-time quantification of binding of fluorescently labeled biomolecules or bacterial cells to immobilized cells or receptors.

Contact:

BICU affinity user club:

Description of the user clubs

  • To become members of the BICU affinity userclub, the PI of your group will have to read the BICU policy and sign a userclub agreement in order to gain access to the instruments within the platform. The agreement is then valid until the end of the year, regardless if it was signed in January or in  December.
  • All active members will recieve a new contract by the end of each year, with the option of signing for another year, or if they would like to pause their membership. The membership itself is free of charge, but introductions to the instruments and booked running time will be invoiced twice a year. 
  • The price on the machines are based on the annual service costs and the amount of users, so prices may change from year to year based on usage. BICU is a non-profit and all the earnings are meant to go back into the instruments in the form of upgrades, softwares , annual services and repairs.
  • You can read more about the individual costs of the instruments below in their respective sections.

How to become a member?

1. Write an email to johan.olofsson.edlund@umu.se to recieve the contract to sign.
2. We will then send a copy of the BICU policy to read and a user contract to be signed by the PI of the group applying.
3. Once the agreement is signed by the PI and sent back to us you are free to start booking introductions.

Introductions and access:

  1. Before using any instrument within the BICU platform you will have to aquire a ”drivers license” for each of the machines that you want to use.
  2. The introductions are given to every new user regardless of previous experience. This is to make sure everyone has the minimum knowledge required to run the instrument properly and in a safe manner for both you and the machine.
  3. We allow a maximum number of 2 people per introduction. This is partly due to the limited space where the instruments are located, but mainly to ensure that everyone gets enough "hands-on" training.
  4. Depending on what instrument you are interested in there may be 1 or 2 people able to sign off on your ”drivers license”. You can always write to johan.olofsson.edlund@umu.se to find out.
  5. Once you have agreed on a time for the introduction, please respect the time of the person giving the introdcution as they all have their own labwork to attend to.
  6. Once the introduction is completed, send your casID to the person that gave you the introduction and they will grant you access to the booking system. If needed they will also give you access to the rooms and corridors where the instrument is located.
  7. Book the instrument as needed and remeber to sign the logbooks on site when using them.

Technical questions & experimental support:

  1. Our staff will be able to help you with getting started and setting up the basic experiments for your first run if needed.
    Just make sure to schedule with one of the contact persons for that instrument a while before you go since they all have other obligations outside of BICU as well.
  2. For more advanced help with experimental assays and data analysis we will do what we can in order to answer your questions and help you out.  We might not be able to answer everything, but depending on what questions you have we might be able to refer you to other users working on similar things, good publications on the topic that we have saved up during the year or some outside expertise that we have connections to.

When errors occur hard/software issues:

  1. Don't panic, some of the machines are a bit old and run on older computers that might loose connection once in a while.
  2. Take a picture if you see an error message or something strange.
  3. Contact  via email at johan.olofsson.edlund@umu.se (or the other responsible for the machine) or call +46738010659 and describe the problem.
  4. Most often it will be minor things that can be handled within a short time. Some errors are reoccuring and can be solved over the phone as well in case you have valuable samples.
  5. Whatever you do, DO NOT tamper with the machine yourselves. Doing so will void eventual service contracts and you/your group might become financially responsible.

Affinity Measurements Instruments

Biacore 3000

More Details

  • Location:
    The room is: Building KBC, 6th floor, room A5 39 07.
    Directions using digital map
  • Price:
    • 1650 SEK/day
    • Introduction/driver license Cost is 1500 SEK (2x half days)
  • About:

    Protein – Protein
    Protein – DNA
    Protein – Small molecule or fragment.
    Protein – Carbohydrate
    Protein – Lipid or Membrane / Liposome interactions
    Protein – RNA

    By attaching one of your targets (the ligand) onto a chip surface and then flowing over your other target (analyte) the biacores optical sensors measures the change in mass bound to the chip. Various commercial chips allow you to attach your ligands in different ways depending on your material. The most common ones are through His-tags, Amine coupling or Biotin-tags.  

    With the kinetic assays you can determine association, dissociation and binding constants for your interactions. This older version of the Biacore gives the user the option of doing “manual” injections to better optimize their running conditions and also create their own scripts for reproducible workflows. For the less experienced one you can also use the application wizard templates and be guided through your experiments step by step with the internal software.

Proteon XPR36

More Details

  • Location:
    The room is: Building KBC, 6th floor, room A5 39 07.
    Directions using digital map
  • Price:
    • 500 SEK/day
    • Introduction/driver license Cost is 1500 SEK (2x half days)
  • 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.

Auto-ITC200

More Details

  • Location:
    The room is: Building KBC, 6th floor, room A5 39 07.
    Directions using digital map
  • 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.

LigandTracer® Green

More Details

  • Location:
    The room is: Building KBC, 6th floor, room A6 31 01.
    Directions using digital map
  • 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.

J-1700 CD Spectrometer

More Details

  • Documentation: J-1700 CD Spectrometer
  • Location: Room A52907
    Directions using digital map
  • Price:
    • Full 24 hours – 800kr
    • 8 hours during the day – 600kr
    • 6 hours during the day – 500kr
    • 4 hours during the day – 400kr
    • 2 hours during the day – 300kr
    • Odd hours 18-08 - 300kr/booking
    • Introduction/drivers license (2hours) - 1000kr
  • About:
    The CD is a form of light absorbtion spectroscopy that measures the the difference in absorbance of right- and left-circulary polarized light. The machine is most commonly used for protein conformations and structural analysis of biomolecules. 
  • Some features of the CD: 
    • Wavelength range 163 - 2500nm
    • Simulatnious Multi-Probe (SMP): CD, LD, flourescense, UV-visible, absorbance
    • Three scanning modes: auto, step and continuous

The CD has also been installed with a karusell for 6 cuvettes to allow measurements 6 samples at a time.  
It also comes with a thermoregulation module that allows you to perform temperature gradient measurments. 

Quartz Crystal Microbalance with Dissipation monitoring (QCM-D)

More Details

  • Location:
    The room is: Building 6M, 4th floor, room 6070.
    6070, 6M, Norrlands universitetssjukhus Nus - MazeMap
  • Price:
    500 SEK/day
    Introduction/driver license Cost is 1500 SEK
  • Contact Person:
    Hudson Pace (hudson.pace@umu.se)
  • 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.
    More Information can be found here: https://awsensors.com/quartz-crystal-microbalance/
Latest update: 2023-09-11