Electron Paramagnetic Resonance Spectroscopy (EPR) services as well as other detection methologies for measuring free radicals, singlet oxygen, nitric oxide and the array of related oxidants and oxidative damage products are available. The EPR facility assists users in the detection of: free radicals in systems that range from solids, solutions, cells, tissues and whole animals, nitric oxide and related metabolites including peroxynitrite, MDA/TBARS, indicators of lipid peroxidation that are detected with fluorescence spectroscopy, oxidation of fluorescent dyes as indicators of oxidation reactions in living cells, UV-Vis spectroscopy for kinetic studies of PhGPx, Cellular oxygen consumption for studying metabolic oxidations reactions, oxidative stress-indicators using HPLC: including DNA damage products, antioxidants such as vitamins C and E, beta-carotene, precise cell volume measurements.
EPR Hours and Contact Information: Services are available 8AM-5PM Monday through Friday (and at other times based on specific discussion with Drs. Spitz and Buettner). To set up an appointment, or if you have questions, please contact: Brett Wagner at brett-wagner@uiowa.edu.
ESR Facility Rates (Effective 08-01-2023)
Rates for University of Iowa users
Small routine supplies necessary for “in facility” sample handing and preparation are included in the above.
Specialty chemicals and supplies are charged at cost, which includes the labor cost for preparation to be user-ready. Because this is a pass-through expense, discounts are not normally available.
For specific assays, such as vitamin C, vitamin E, or superoxide, charges are calculated by hour of instrument use plus appropriate analysis and labor costs. A 30% discount for the analysis of vitamin C in patient samples for UIHC/HCCC clinical trials is provided.
Rates are based on anticipated costs for repair and upkeep, labor involved in calibration, quality control, and the cost of small supplies.
A 30% discount is provided for instrument use by HCCC/CCOM members.
| ESR Instrument use (Standard configuration | $84 h-1 | HCCC $60 h-1 |
| ESR Instrument use (Low temperature) | $104 h-1 | HCCC $72 h-1 |
| Nitric Oxide Analyze Set-up fee | $60 h-1 $55 h-1 | HCCC $44 h-1 HCCC $39 h-1 |
| Beckman LS-50-B Fluorescence | $50 h-1 | HCCC $28 h-1 |
| Oxygen Monitor systems (YSI or Biostat) | $16 h-1 | HCCC $12 h-1 |
| Seahorse Instrument | $136 h-1 | HCCC $96 h-1 |
| Supplies: Consumables | $132 (typical costs of plate and media) | |
| Mito reagents | $17 each (*Typical Seahorse Experiments require 1.5-2.0 h instrument time) | |
| Labor charges | $100 h-1 | HCCC $72 h-1 |
3) Antioxidant Enzyme Services (AES)
Services:
The AES will carry out analysis of major antioxidant enzymes (i.e., CuZnSOD, MnSOD, EcSOD, CAT, GSTs, GGTs, GCS, and GPx), O2·-, H2O2, and soluble small molecular weight thiols/redox couples [i.e., cysteine, g-glutamylcysteine, homocysteine, N- acetylcysteine, GSH/GSSG, NADPH/NADP+, Trx(SH)2/TrxS2] and electron transport chain proteins as well as:
- Measure electron transport chain complex (I-V) activities with protein assays for normalization 40$/sample
- Measure thiols/redox couples and antioxidant enzyme activities.
CuZnSOD, MnSOD, EcSOD, CAT, GSTs, GGTs, GSH/GSSG, NADPH/NADP+, and GPx activities with protein assay for normalization 40$/sample
- Measure immunoreactive protein for antioxidant enzymes.
Cost of antibodies, supplies, and personnel (45$/hour)
- Determine steady state levels of mRNA for redox biology proteins with quantitative PCR; Cost of primers, supplies, and personnel (45$/hour)
- Transfect antioxidant enzyme sense and antisense cDNAs into user specified cell lines and characterize the alterations. Cost of supplies and personnel (45$/hour)
- Maintain, develop, and provide reagents for the study of antioxidant/ pro-oxidant proteins and O2·-, H2O2 measurements in living cells and tissues: i.e., fluorescent probes, antibodies, cDNAs, vectors, and cell lines; Cost of supplies and personnel (45$/hour)
- Antisense/siRNA/adenoviral/lentiviral reagents for antioxidant proteins; Cost of supplies and personnel (45$/hour)
- Provide O2 environments at varying levels from 1% to 20% O2. Cost of supplies and personnel (45$/hour)
Rates: The regular charges for non-UI members associated with the AES are negotiated with each investigator and standard guidelines for charges determined as outlined above; with the basic principle that the core wants to facilitate the user’s science in the most cost-effective way that generates the most impactful results while breaking even financially.
If the core lab staff runs the experiment the cost of personnel is included. If the investigator provides the personnel to be trained by the facility, the cost of core personnel is reduced to that required to be provided by core lab staff under each specific circumstance. The investigators are also responsible for the added specific supplies cost unique to their experiment. The charges are discounted 30% for CCOM/HCCC members or users directly collaborating with Iowa CCOM/HCCC members. Outside users are charged the full rate.
AES Hours and Contact Information: Services are available 8AM-5PM Monday through Friday (and at other times based on specific discussion with Drs. Spitz). To set up an appointment, or if you have questions, please contact Douglas Spitz at douglas-spitz@uiowa.edu.