Highly sensitive protein detection based on lanthanide chelates with antenna ligands providing a linear range of five orders of magnitude.
Protein detection is an important task for pharmaceutical and clinical research today. Numerous protein staining techniques exist but are limited regarding their sensitivity and often narrow linear quantification ranges. To the best of our knowledge, this is the first description of a novel class of lanthanide chelatators, which absorb in the lower energy region at 360 nm. The new compound (6,9-dicarboxymethyl-3-{4-([1,10]-phenanthrol-2-ylethinylphenyl-carbamoyl)-methyl}-3,6,9-triaza-)-undeca-1,11-dicarboxylic acid) was coupled to different proteins and showed highly sensitive protein detection limits in both sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (1.5 fmol of bovine serum albumin) as well as Dot Blot (100 amol of lysozyme). Furthermore, the novel compound shows an exceptionally broad linear quantification range over 5 orders of magnitude allowing applications that require the highest sensitivity alongside standard sensitivity. In addition, the new compound offers multiplexing capabilities.
Zuchner
Thole
T
Institute of Bioanalytical Chemistry, Center of Biotechnology and Biomedicine, Faculty of Chemistry and Mineralogy, Leipzig University, Deutscher Platz 5, 04103 Leipzig, Germany. zuechner@rz.uni-leipzig.de
Schumer
Frank
F
Berger-Hoffmann
Renate
R
Müller
Katrin
K
Lukas
Mathias
M
Zeckert
Kornelia
K
Marx
Jörg
J
Hennig
Horst
H
Hoffmann
Ralf
R
D002614:Chelating Agents / Q000737:chemistry*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D006801:Humans; D028581:Lanthanoid Series Elements / Q000737:chemistry*; D011506:Proteins / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
D002614:Chelating Agents; D028581:Lanthanoid Series Elements; D011506:Proteins