TRAINING SCHOOL ON METALLOTHIONEIN
Title: Metallothionein and its relation to zinc (II) ions
Where: Zemedelska 1, Brno, Czech Republic
(http://web2.mendelu.cz/af_239_nanotech/04.php?show=4)
Date: 15th - 17th June 2015 (8 h/day)
Content: Laboratory expertise in multi-instrumental analysis of metallothionein and zinc (II) ions.
Requirements: Post docs and PhD students.
The participants will be divided into groups of 3 or 4 persons
Practical sub-courses:
1. Determination of metallothionein using electrochemistry. Hynek
2. Electrophoresis and immunodetection. Krizkova
3. Analysis of metallothionein expression at nucleic acids level. Masarik / Gumulec
4. Immunohistochemistry. Kominkova / Michalek
5. Matrix assisted laser desorption/ionization. Merlos Rodrigo
Maximum participants: 9 or 12
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The Network for the Biology of Zinc (Zinc-Net) TD1304
TRAINING SCHOOL ON METALLOTHIONEIN
Day 15th June
10:00
Summer School Opening
prof. RNDr. Ladislav Havel, CSc. - Rector of Mendel University in Brno
Invited lectures
10:20
Zinc signals and the control of immune cell functions
Hajo Haase (Aachen University, Germany)
11:00
Metallothionein and neuroinflammation
Juan Hidalgo (Autonomous University of Barcelona, Spain)
Lunch
Plenary lectures
13:00
Metallothionein as cancer biomarker
Jaromír Gumulec
13:30
Metallothionein as zinc protein
Michal Masarik
14:00
Metallothionein and oxidative stress
Branislav Ruttkay-Nedecky
The overview of the presentations will be published in Journal of Metallomics and
Nanotechnologies
14:30 – 15:00
Coffee Beak
15:00-17:00
LABORATORY OF METALLOMICS AND NANOTECHNOLOGIES LABTOUR
17:30
GUIDED BRNO BUS TOUR - Vlastimil Sochor
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The Network for the Biology of Zinc (Zinc-Net) TD1304
TRAINING SCHOOL ON METALLOTHIONEIN
16th and 17th June
List of experiments
Electrochemical detection of MT (David Hynek)
1) Preparation of sample
2) MT determination
3) MT determination coupled with Adsorptive transfer technique
4) Automated determination
5) Evaluation and data processing
Metal chelate affinity chromatography (Vladimir Pekarik)
1) Bacterial transformation
2) Induction of protein expression
3) Protein isolation – protein fractionation (acetone precipitation)
4) Test affinity of MT for different metals – NTA (nitrilotriacetic) agarose Cd, Ni, Cu, Fe, Zn,
empty. Comparison with PrP control Elution –EDTA
5) Detection: electrochemistry, Western-Blot, chemical staining with Fl-AsHor Re-AsH
(depending on Pavel ability to synthetize the compounds).
Capillary electrophoresis (Marketa Vaculovicova)
1) in vitro MT labelling by fluorescent dye
2) CE-LIF analysis of fluorescent MT
3) CE-UV analysis of MT clones
4) Data analysis
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The Network for the Biology of Zinc (Zinc-Net) TD1304
TRAINING SCHOOL ON METALLOTHIONEIN
ELISA I (Sona Krizkova)
1) Samples and standards preparation
2) Incubation
3) Well surface blocking
4) Washing
5) Application of primary antibody
ELISA II (Petr Michalek)
1) Incubation with secondary antibody
2) Application of chromogenic substrate
3) Absorbance measurement and data analysis
Spectrophotometry + chip CE (Sona Krizkova)
1) ApoMT interaction with Zn
2) Incubation and spectrophotometric detection
3) Chip electrophoresis
All experiments will be performed according to the protocols (Nature Protocols format)
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The Network for the Biology of Zinc (Zinc-Net) TD1304
TRAINING SCHOOL ON METALLOTHIONEIN
Schedule:
10:00
Summer School Opening
10:20-12:00
INVITED LECTURES
12:00-13:00
LUNCH
13:00-14:30
PLENARY LECTURES
15:00-17:00
LABTOUR
17:30
GUIDED BRNO BUS TOUR
15th June
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The Network for the Biology of Zinc (Zinc-Net) TD1304
TRAINING SCHOOL ON METALLOTHIONEIN
9:00-11:00
11:00-13:00
13:00-14:00
Elisa I (Krizkova)
Metal chelate affinity
Electrochemistry of MT
chromatography (Pekarik)
(Hynek)
Electrochemistry of MT
Metal chelate affinity
CE (Vaculovicova)
(Hynek)
chromatography (Pekarik)
LUNCH
LUNCH
LUNCH
Elisa II (Michalek)
Elisa I (Krizkova)
Metal chelate affinity
16th June
14:00-16:00
16:00-18:00
9:00-11:00
11:00-13:00
17th June
13:00-14:00
14:00-16:00
chromatography (Pekarik)
Spectro + chip CE
Electrochemistry of MT
Metal chelate affinity
(Krizkova)
(Hynek)
chromatography (Pekarik)
Dinner Party
Stopkova Plzeňská Pivnice
Metal chelate affinity
Elisa II (Michalek)
Elisa I (Krizkova)
CE (Vaculovicova)
Spectro + chip CE
chromatography (Pekarik)
Metal chelate affinity
chromatography (Pekarik)
(Krizkova)
LUNCH
LUNCH
LUNCH
CE (Vaculovicova)
Spectro + chip CE
Elisa II (Michalek)
16:30
(Krizkova)
Summer School Closing
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The Network for the Biology of Zinc (Zinc-Net) TD1304
TRAINING SCHOOL ON METALLOTHIONEIN
Accommodation: it is possible to accommodate participant on very near situated dormitory or
in the hotels nearby.
Avanti Hotel Střední 61, Brno 4
Hotel Bobycentrum Brno, Sportovní 559/2A, Brno 3
Bird's Nest, Tišnovská 94, Brno, 3
Villa JKC, Lieberzeitova 20, Brno, 3
Point Pension-Restaurant, Vranovská 58, Brno 3
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The Network for the Biology of Zinc (Zinc-Net) TD1304