Download Lab Spotlight on Odyssey Expert: Dr. Andreas - LI

Lab Spotlight
Exceptional Researchers and LI-COR Imaging Systems Driving Results
Spotlight on Dr. Andreas von Knethen
Dr. Andreas von Knethen
is a Postdoctoral Fellow in the Institute of
Biochemistry at Johann
Wolfgang Goethe-University in Frankfurt, Germany.
His research focuses on
immune paralysis, tumor “Not only are the
biology, inflammation, and
EMSAs easier, behypoxia.
cent electrophoretic mobility shift
One of the main focuses in Dr von
Quantitative Western blots are also
Knethen’s lab is apoptosis of T cells.
“In sepsis, you have two phases. The
first one is the hyper-inflammatory
phase, which is characterized by the
release of pro-inflammatory cytokines
and other pro-inflammatory markers.
The second is the hypo-inflammatory phase. This phase is also called
immune paralysis because the immune-competent cells are decreased
cause you don’t have
to radioactively label
anything, but also the
Western blots can be
saved for years because
there is no chemiluminescence that is
decreasing after thirty
minutes.”
- Dr. Andreas von Knethen
You just run the gel and put it onto
the Odyssey, and you can see the
DNA binding of the proteins. You
don’t need to dry the gel and there is
no radioactivity.”
useful. Dr. von Knethen uses the
Odyssey to analyze protein phosphorylation with two-color Westerns.
“You can use the red and green colors together to see the total amount
of protein and the phosphorylation,
to analyze both of them on the same
blot.” For gene knockdown studies,
he uses Odyssey Westerns to monitor protein levels. He considers it
the mRNA is down-regulated.”
sepsis are associated with T cell apop-
cell apoptosis during sepsis. Fluores-
makes the assays “much more easy.
protein is really gone, and not only
leading to immune paralysis during
transcription factor responsible for T
with IRDye® 700 or IRDye 800, which
the Western blot just to prove that the
second or new infection. Mechanisms
LI-COR Odyssey® to analyze PPARγ, a
nucleotides are fluorescently labeled
at the protein level. “You have to do
not able to answer adequately to a
Dr. von Knethen’s group uses the
binding of PPARγ to DNA. The oligo-
very important to confirm knockdown
or inactivated and then the body is
tosis, and lead to depletion of T cells.”
assays (EMSA) are used to examine
LINKS
For more information about Dr. von Knethen’s research, visit the link below:
http://www.pathobiochemie1.de/research/index.php
He feels that the Odyssey makes things easier in the
We thank Dr von Knethen for his contributions in bio-
lab. “Not only are the EMSAs easier, because you
medical research, and are proud to consider him an
Odyssey Expert.
don’t have to radioactively label anything, but also the
Western blots can be saved for years because there is
no chemiluminescence that is decreasing after thirty
minutes.”
PUBLICATIONS
Publications resulting from work on the Odyssey:
1.
von Knethen A, Neb H, Morbitzer V, Schmidt MV, Kuhn AM, Kuchler L, Brüne B. (2011) PPARgamma stabilizes HO-1 mRNA in
monocytes/ macrophages which affects IFN-beta expression. Free Rad Biol Med, 51:396-405.
2.
Kuhn AM, Tzieply N, Schmidt MV, von Knethen A, Namgaladze D, Yamamoto M, Brüne B. (2011) Antioxidant signaling via Nrf2
counteracts lipopolysaccharide-mediated inflammatory responses in foam cell macrophages. Free Rad Biol Med, 50:1382-1391.
3.
Schmidt MV, Paulus P, Kuhn AM, Weigert A, Morbitzer V, Zacharowski K, Kempf V, Brüne B, von Knethen A. (2011) PPARgammainduced T-cell apoptosis reduces survival during polymicrobial sepsis. Am J Res Crit Care Med, 184:64-74.
4.
Barra V, Kuhn AM, von Knethen A, Weigert A, Brüne B. (2011) Apoptotic cell-derived factors induce arginase II expression in
murine macrophages by activating ERK5/CREB. Cell Mol Life Sci, 68:1815-1817.
5.
Weigert A, Cremer S, Schmidt MV, vvon Knethen A, Angioni C, Geisslinger G, Brüne B (2010) Cleavage of sphingosine kinase 2
by caspase-1 provokes its release from apoptotic cells. Blood, 115:3531-3540.
6.
Jennewein C, von Knethen A, Schmid T, Brüne B (2010) MicroRNA-27b contributes to lipopolysaccharide-mediated peroxisome
proliferator-activated receptor gamma (PPARgamma) mRNA de-stabilization. J Biol Chem, 285:11846-11853.
7.
Namgaladze D, Morbitzer D, von Knethen A, Brüne B (2010) Phospholipase A2-modified low-density lipoprotein activates macrophages peroxisome proliferators-activated receptors. Arterioscler Thromb Vasc Biol, 30:313-320.
8.
von Knethen A, Tzieply N, Jennewein C, Brüne B (2010) Casein kinase-II-dependent phosphorylation of PPARgamma at Ser16
and Ser21 provokes CRM1-mediated shuttling of PPARgamma from the nucleus to the cytosol. J Cell Sci, 163:192-201.
9.
Namgaladze D, Jennewein C, Preiss S, von Knethen A, Brüne B (2009) Attenuated suppression of the oxidative burst by cells
dying in the presence of oxidized low density lipoprotein. J Lipid Res, 50:2173-81.
10. Weis N, Weigert A, von Knethen A, Brüne B (2009) Heme oxygenase-1 contributes to an alternative macrophage activation
profile induced by apoptotic cell supernatants. Mol Biol Cell, 20:1280-8.
11. Jennewein C, Kuhn AM, Schmidt MV, Meilladec-Jullig V, von Knethen A, Gonzalez FJ, Brüne B (2008) Sumoylation of PPARgamma by apoptotic cells prevents LPS-induced NCoR removal from kappaB binding sites mediating transrepression of proinflammatory cytokines. J Immunol, 181:5646-52.
12. von Knethen A, Soller M, Tzieply N, Weigert A, Johann AM, Köhl R, Brüne B (2007) PPARgamma attenuates cytosol to membrane translocation of PKCalpha to desensitize monocytes/ macrophages. J Cell Biol, 176:681-694.
4647 Superior St. • P.O. Box 4000 • Lincoln, Nebraska 68504 USA
North America: 800-645-4267 • International: 402-467-0700 • FAX: 402-467-0819
LI-COR GmbH (Germany, Austria, Switzerland, Czech Republic, Hungary, and Slovakia):
+49 (0) 6172 17 17 771 • LI-COR UK Ltd.: +44 (0) 1223 422104 • www.licor.com
LI-COR
Odyssey
trademarks
or
LI-COR is
is an
an ISO
ISO 9001
9001 registered
registeredcompany.
company. ©
© 2011
2007LI-COR
LI-CORInc.
Inc.Specifications
Specificationssubject
subjectto
tochange.
change.LI-COR,
LI-COR,IRDye,
Aerius,and
Odyssey
andare
IRDye
are registered
trademarks of
registered
trademarks
of LI-COR, Inc.
All other
trademarks
toThe
theirAerius
respective
owners.
LI-COR, Inc.
All other trademarks
belong
to their
respecivebelong
owners.
Automated
Infrared Imager, Odyssey Infrared Imager, and IRDye reagent
products are covered by U.S. patents, foreign equivalents, and other patents pending.
0711
1207 982-12318
982-09493