The Core Facility's Proteomics/Biochemistry Resources include a range of techniques from Western blot/DNA gel imaging, NanoTemper's MST technology to measure molecular interaction in solution, EliSpot/FluoroSpot analysis, Multiplexing analysis technology to column chromatography and more.
Available Instruments
Bio-Techne Jess Automated Western Blot System
The Jess Western Blot system is an automated capillary-based immunoassay instrument capable of automatically running up to 25 samples for fluorescence, chemiluminescence or protein normalization imaging, quantification and analysis assays.
Applications
The Jess Western Blot system is an automated, capillary-based immunoassay system that integrates protein separation, immunoprobing, detection and analysis into one instrument. Samples and reagents are simply loaded onto a microplate while the system performs the steps of Western blotting in a precise and reproducible way. The system can also automatically run a second immunoassay on the same sample/capillaries if required. The system allows for fluorescent detection, chemiluminescent detection, protein normalization and even classic blot imaging if necessary with software driven analysis and quantification of results.
Specifications
Sample required: Total protein: 0.3-1.2 µg. Chemiluminescence: 0.6-1.2 µg. Fluorescence: 2-4 µg. Protein Normalization: 0.6-4 µg.
Volume required: 3µl/well
Size Range: Molecular weight ladder ranges from 2-440 kDa
Resolution: ±15-20% for MW <20 kDa and ±10% for MW >20 kDa.
Dynamic Range: Total protein: 2-3 logs. Chemiluminescence: 3-4 logs. Fluorescence: 4 logs. Protein Normalization: 1 log.
Sensitivity: Total protein: ng. Chemiluminescence: low pg. Fluorescence: low pg. Protein Normalization: ng.
Run Time: 3 hours
Samples per run: 13 or 25
Software: Compass for experimental protocol and sample analysis.
User Information
For more information on the Jess Western blot system, visit:
https://www.proteinsimple.com/jess.html
Azure c500 Imaging System
The Azure c500 is a multi-faceted imaging system for Western blots and DNA gels and includes AzureSpot, a separate analysis software for quantification.
Applications
The c500 is designed to image DNA gels and Western Blots using both chemiluminescence or Near-InfraRed fluorescence (allowing for two color imaging on the same blot). The system also comes with AzureSpot Analysis Software for quantitative analysis.
Specifications
Lasers at 660 nm and 785 nm: With a narrower excitation band than LEDs or white light sources, you get NIR imaging with better sensitivity and lower background.
Epi blue LED at 470 nm: Image blue-excited DNA dyes like SYBR® Safe, a feature available on all cSeries models.
Epi White Lights: Uniform overhead illumination for white light imaging.
Trans White Imaging: Image visible/white light dyes, such as Coomassie Blue, in gels or other translucent samples.
Dual-Wavelength UV at 302 nm and 365 nm: Image DNA gels and more with a UV light source compatible with a wide range of dyes, including ethidium bromide, SYBR® green, SYBR gold, SYPRO® orange, fluorescein, RadiantRed®, TexasRed®, and SYPRO Red.
Touch-Screen Controls: Easily manage data acquisition and analysis with the touch of a finger, driven by Windows OS.
High Resolution Camera: Capture fine details of your sample.
Deep Peltier Cooling: Experience excellent image quality and reduced noise with low temperature camera cooling (-50°C).
Dual-Focus Technology: Get perfectly-focused images and optimal lane settings without having to touch the camera.
7-Position Filter Wheel: Perform a wide range of applications with a motorized, multi-position filter wheel.
Chem Blot Shelf: Get better sensitivity by placing chemiluminescent blots closer to the detector. The adjustable shelf can be stored in the door when not in use.
User Information
For more information on the c500 imaging system, visit:
https://www.azurebiosystems.com/cseries-imaging-systems-old/
https://www.azurebiosystems.com/wp-content/uploads/2016/06/cSeries-brochure_24.pdf
AID vSpot Spectrum
The vSpot Spectrum is an advanced EliSpot/FluoroSpot imaging and analysis system designed for a variety of applications and plate formats.
Applications
The vSpot Spectrum is both an Elispot and Fluorospot reader and is also capable of analyzing a variety of assays like viral plaque assays, neutralization assays and colony counting. Plate formats can vary from 6 to 384-well depending on application.
Specifications
Light sources: LED ring illumination for color imaging and additional LED SOLA SMII light source from Lumencor for fluorescence.
Filters: 3 narrow-band hard coated fluorescent filters on board for FITC, Cy3 and Cy5.
Camera: 2 digital firewire color cameras, 5 megapixels each, optimized for fluorescence
Software: The vSpot Spectrum is powered by AID EliSpot 7.0 and 8.0 software for acquisition and analysis.
User Information
For more information on the vSpot Spectrum system, visit:
https://www.aid-diagnostika.com/en/reader-systems/aid-vspot-spectrum
For information on available kits from AID:
https://www.aid-diagnostika.com/en/kits/elispot-assay/t-cell-assay/basic-kits
NanoTemper Monolith NT.115
The Monolith allows for rapid and simple molecular interaction measurements (Kd) in solution and in small sample volume using MicroScale Thermophoresis technology.
Applications
The Nanotemper Monolith system is designed to measure molecular interactions (Kd) within minutes. Contrary to better known SPR systems with costly consumables and complex immobilization requirements, the Monolith allows measurement of interactions in solution (cell lysates) and small volumes, facilitating sample preparation, staining and scope of experimentation.
Specifications/Methods
MicroScale ThermophoresisTM (MST) provides an easy, fast and precise approach to quantify biomolecular interactions. It allows for the detection of a wide range of interaction types, ranging from ion and fragment binding up to interactions of macromolecular complexes such as liposomes or ribosomes. The main key feature of MST is the capability to work under close-to-native conditions: Measurements are performed immobilization-free and in any buffer, or even in complex bioliquids such as serum and cell lysate. In addition, the method combines straight forward sample preparation, low sample consumption of less than 4 μl per sample at nM concentration, and a large dynamic range with dissociation constants from the sub-nM to mM range. The NanoTemper Technologies Monolith® instruments are maintenance-free, have low running costs and use an intuitive software user interface with provides guided assay development and automated data analysis, making MST a one-of-a-kind, low-hurdle biophysical technology for every lab.
User Information
For more information on the Monolith NT.115 system, visit:
https://nanotempertech.com/monolith/monolith-nt115/
For information on consumables and staining kits from Nanotemper, visit :
https://nanotempertech.com/monolith-consumables
AKTA Pure Protein Purification System
The AKTA Pure system in a flexible column chromatography system with advanced setting capabilities for a broad range of applications.
Applications
The ÄKTA Pure is a flexible chromatography system for purification of proteins, peptides, and nucleic acids from microgram to gram levels of target product,
in various research applications. It allows for quick and easy customization thanks to the modular design of both the hardware and software. The ÄKTA pure 25 has a flow rate up to 25 mL/min and is equipped with a F9-R Fraction collector.
Specifications/Methods
Among the applications supported are affinity chromatography, size exclusion chromatography (SEC, also known as gel filtration), ion exchange chromatography, hydrophobic interaction chromatography, and reversed phase chromatography (RPC).
User Information
For information on the AKTA Pure system, visit this site.
Resources for techniques and troubleshooting can be found here.
Consumables are available here and here.
AKTA Start Protein Purification System
The AKTA Start is a simpler chromatography system for laboratory scale protein purification.
Description
AKTA start is a preparative chromatography system for laboratory scale protein purification. it is designed as a stand-alone system to purify a wide variety of proteins using built-in quick start methods or predefined templates, or by creating your own methods.
Combined with the Frac30 fraction collector, it allows automated fraction collection in volumes from 0.5 to 15 mL.
User Information
For more information and resources on the ÄKTA Start purification system, visit this page.
Compatible prepacked columns are available here.
SpectraMax iD3 Plate Reader
The SpectraMax iD3 is a multi-user microplate reader for a wide range of application using absorbance, fluorescence or luminescence.
Applications
The SpectraMax iD3 is a multi-user microplate reader capable of measuring absorbance, fluorescence and luminescence. Possible applications include cell viability assays, DNA/RNA quantitation, Reporter gene assays, etc..
Specifications
Wavelength ranges :
- Absorbance: 230 - 1000 nm
- Fluorescence Excitation: 250 - 830 nm
- Fluorescence Emission: 270 - 850 nm
- Luminescence: 300 - 850 nm
Absorbance photometric accuracy/linearity: < ±0.010 OD ±1.0%, 0 - 3 OD VIS 0 - 3 OD UV
Absorbance photometric precision/repeatability: < ±0.003 OD ±1.0%, 0 - 3 OD VIS 0 - 3 OD UV
Sensitivity top read optimized: FL: 1 pM fluoresein 96 well, 1 pM fluorescein 384 well Lumi: 2 pM ATP 96 well, 4 pM.
Light Source: High power xenon flash lamp
Detector(s): Silicon photodiode, Ultra-cooled photomultiplier tube
Read times (kinetic interval for 96-well plate)*: Abs: 28 seconds, FL: 21 seconds,Lumi: 21 seconds
Plate shaking: Linear, Orbital, Double orbital
Plate type(s): 6-384 well plates.
User Information
For more information on the SpectraMax iD3, visit this page.
MAGPIX Luminex System
The MAGPIX Luminex System is designed for multiplexing analysis of a large number of analytes for a wide range of applications through commercial or custom designed kits and solutions.
Applications
The MAGPIX Luminex system allows for multiplexing analysis of up to 50 analytes using Luminex® xMAP® technology with a wide range of research applications. Commercial kits and solutions for protein and RNA detection assays compatible with the MAGPIX system are available for various manufacturers.
Description
Microsphere beads, either polystyrene or paramagnetic, are color-coded into up to 500 distinct sets. Each bead set can be coated with a reagent specific to a particular bioassay, allowing the capture and detection of specific analytes from a sample. Inside the Luminex® analyzer, a light source excites the internal dyes that identify each microsphere particle, and also any reporter dye captured during the assay. Multiple readings are made on each bead set, which further validates the results. Using this process, xMAP® technology allows multiplexing of up to 500 unique bioassays within a single sample, both rapidly and precisely.
User Information
To find kits specific to your research topics and animal models, visit the Luminex Kit Finder site.
For information on the xMAP multiplexing technology, visit this site.
For more information on the MAGPIX system, visit the instrument's overview page.
For resources on use, troubleshooting and experiment setup, check out their instrument resources page.
