Proteomics and Metabolomics Core Virtual Tour Video
Nebraska Center for Biotechnology
Author
06/02/2021
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Description
Virtual tour of the Proteomics and Metabolomics Core Facility. The Proteomics and Metabolomics Core provides high quality service and collaborative work in the fields of Proteomics and Metabolomics to support scientists and students within our University community and around the world.
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- [00:00:00.679](upbeat modern music)
- [00:00:07.650]My name is Sophie Alvarez
- [00:00:09.530]and I'm the director of the Proteomics
- [00:00:11.500]and Metabolomics Facility
- [00:00:13.060]which is part of the Nebraska center for biotechnology.
- [00:00:16.570]Our facility is located on the first floor
- [00:00:18.716]of the Beadle Center.
- [00:00:20.670]The facility was established back in 2015
- [00:00:24.040]to meet an ever-increasing need
- [00:00:25.830]for a range of technical services using mass spectrometry.
- [00:00:29.430]These services include identification
- [00:00:31.860]and relative quantification of proteomics
- [00:00:34.430]and several advanced methods for profiling
- [00:00:36.740]and quantitation of small molecules.
- [00:00:39.470]The primary aim of the facility
- [00:00:41.270]is to provide high quality service and expertise
- [00:00:44.140]for faculty, students, and staff at UNL
- [00:00:47.270]in the fields of proteomics and metabolomics.
- [00:00:50.950]To accomplish this goal, the facility is equipped
- [00:00:53.590]with state of the art instruments.
- [00:00:56.330]The proteomics platform includes an Orbitrap Eclipse Tribrid
- [00:00:59.690]Mass Spectrometer coupled to a Dionex U3000 Nano HPLC
- [00:01:04.020]to provide extremely high resolution,
- [00:01:06.130]high mass accuracy data.
- [00:01:08.300]We routinely run comprehensive experiments
- [00:01:10.890]for protein identification and differential proteomics
- [00:01:14.600]using either label-free or labeling approaches.
- [00:01:18.360]The labeling method utilizes the tandem mass tag
- [00:01:21.000]or TMT chemistry, which allows to combine up to 16 samples
- [00:01:24.263]into one run, reducing experimental variability.
- [00:01:28.750]The total number of protein identified
- [00:01:30.980]ranges from 2,000 to 8,000 proteins
- [00:01:34.210]depending on the experiment design.
- [00:01:36.700]This platform is also used to identify proteins
- [00:01:39.390]from cool-down acids in interactions to these
- [00:01:42.290]of single proteins from purification experiments.
- [00:01:46.050]For metabolomics, these needs are addressed
- [00:01:48.380]by the use of multiple instrument capabilities.
- [00:01:51.650]For more basic analysis we use a standalone
- [00:01:54.373]Agilent Ultra-.High Performance Liquid Chromatography System
- [00:01:57.770]or UHPLC which is equipped with a diode-array detector,
- [00:02:01.860]a fluorescence detector and evaporative light
- [00:02:04.500]scattering detector or yellow steel.
- [00:02:07.600]This provides a robust cost effective platform
- [00:02:10.940]to establish targeted assay of a wide range
- [00:02:13.550]of compounds from common molecules like amino acids
- [00:02:17.420]and carbohydrates to more specialized metabolites
- [00:02:20.700]like carotenoid and phospholipids.
- [00:02:23.520]Our gas chromatography mass spectrometer or GCMS
- [00:02:27.220]allows for the detection
- [00:02:28.510]of volatile compounds found in foods
- [00:02:30.600]of all the tissue samples and offers a targeted approach
- [00:02:34.353]to studying polar metabolites,
- [00:02:36.430]such as those involving carbon metabolism.
- [00:02:39.630]For projects requiring increased sensitivity
- [00:02:42.600]when compounds are a very low abundance,
- [00:02:44.950]the facility offers a liquid chromatography MS platform.
- [00:02:49.040]This consists of a 6,500 Qtrap mass spectrometer
- [00:02:52.143]with a Shimadzu UHPLC on the front.
- [00:02:55.540]With this we can very carefully target particular molecules
- [00:02:58.930]using a technique called multiple reaction monitoring
- [00:03:02.690]or MRM for short, example of the MRM assays available
- [00:03:07.100]on this platform include phytohormones, flavonoids,
- [00:03:10.991]alpha and beta acids from hops and by acids.
- [00:03:15.840]All of these three pieces of equipment, the HPLC, GCMS
- [00:03:20.240]and the Qtrap are open access for students, staff
- [00:03:23.900]or faculty to set up their own methodologies
- [00:03:27.100]and training to do this is provided by the facility.
- [00:03:30.920]Finally for LCMS and targeted metabolomics
- [00:03:34.160]we offer our Q Exactive High Field Orbitrap or QEHF.
- [00:03:38.570]This is a high resolution accurate mass system
- [00:03:41.090]coupled to a thermal vanquish UPLC.
- [00:03:44.250]It is set up to run reverse space
- [00:03:46.370]and hydrophilic interaction liquid chromatography
- [00:03:49.310]for increased metabolomic coverage.
- [00:03:52.002]For all the platforms mentioned both proteomic
- [00:03:55.240]and metabolomics the post acquisition data analysis
- [00:03:58.980]is supported by several established pipelines
- [00:04:01.340]of bioinformatic tools run on powerful computing servers
- [00:04:05.200]and workstations, with these specialized platforms
- [00:04:08.740]the core facilities brought impact
- [00:04:10.480]on research includes, advancing world-class research,
- [00:04:14.210]supporting grant proposals, assisting
- [00:04:16.395]in the education of future scientists
- [00:04:19.460]and supporting economic and scientific advancements.
- [00:04:23.080]Our outreach and education program includes workshops
- [00:04:26.520]seminars, and courses.
- [00:04:28.790]We also provide the services to academic labs outside
- [00:04:32.000]of Nebraska and to private industry.
- [00:04:34.910]Full service and self-service options
- [00:04:36.820]are able to meet the need and budget of our users.
- [00:04:40.210]For more information please visit our webpage
- [00:04:43.000]or follow us on Twitter.
- [00:04:44.696](upbeat modern music)
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