The workshop will be limited to 25 participants on a first-come first-served basis.
| 09.00 – 10.30: | Introduction, Professor Malcolm Jackson. |
| 10.30 – 12.30: | Laboratory-based practical sessions. |
| 12.30 – 13.30: | Lunch and informal discussions. |
| 13.30 – 15.30: | Laboratory-based practical sessions. |
| 15.30 – 16.00: | Tea/Coffee |
| 16.00 – 18.00: | Laboratory-based practical sessions. |
| 18.00 – 18.30: | Informal discussions and departure. |
The format of the day will depend upon the numbers of individuals who are registered for the Workshop, but we anticipate that the delegates will split into 3 groups and spend up to 2 hours at each of the stations listed below. Due to Health and Safety restrictions, participants will primarily be observing experienced laboratory workers undertake the methods.
Introduction. General overview by Professor Jackson on the appropriate methods available for detection of ROS activity in cells and tissues, including mitochondria (presented by Dr Van Remmen).
Station 1. Intracellular detection of ROS. Techniques will include microscopic imaging of ROS with DCFH or other reagents and HPLC to detect specific ROS products of DCFH such as DHE and the use of intracellular spin probes in cells and tissues to determine superoxide or nitric oxide activities by EPR.
Station 2. Extracellular detection of ROS. Techniques will include analysis of extracellular superoxide activity in cells and tissues using cytochrome c and the use of extracellular spin probes to determine superoxide or nitric oxide activities by EPR.
Station 3. Measurement of markers of ROS activity and adaptive responses to changes in ROS. This will include detection of protein sulphydryl, carbonyl and glutathione content in cells and tissues. Adaptive responses to ROS will include determination of redox-sensitive transcription factor activation, determination of antioxidant enzyme activities and stress protein content (protein and mRNA using QPCR).
Upon registration, we will ask individuals to register their interests in learning particular techniques. The Programme may be modified accordingly. Suggestions for any techniques which are not currently listed will also be considered at that time.