6
concentration of 20 nM. 10 µl of BLIP-
containing cell lysate was mixed with 10 µl
TEM1 dilutions. For measurements, samples
were filled into standard treated capillaries
(Cat#K002).
Instrumentation
The measurements were conducted on a
NanoTemper Monolith NT.115 instrument.
The Analysis was performed at 50 % LED
power and 80 % MST power, Fluo. Before
time was 5 seconds, MST on time was
30 seconds and "Fluo. After" time was
5 seconds.
.
References
.
Wang et al., Thermodynamic investigation of the
role of contact residues of beta-lactamase-
inhibitory protein for binding to TEM-1 beta-
lactamase. J Biol Chem. 2007 Jun
15;282(24):17676-84.(2007).
Reichmann et al., The modular architecture of
protein-protein binding interfaces. Proc Natl Acad
Sci U S A. Jan 4;102(1):57-62- (2005)
Jerabek-Willemsen et al., Molecular interaction
studies using microscale thermophoresis. Assay
Drug Dev Technol. Aug;9(4):342-53 (2011).
Wienken et al., Protein-binding assays in
biological liquids using microscale
thermophoresis. Nat Commun. Oct 19;1:100
(2010).
Albeck S and Schreiber G, Biophysical
characterization of the interaction of the b-
lactamase TEM-1 with its protein inhibitor BLIP,
Biochemistry 38, 11–21 (1999)
Philipp et al., Protein-binding dynamics imaged in
a living cell.
Proc Natl Acad Sci U S A. Jan 31;109(5):1461-6
(2012).
© 2012 NanoTemper Technologies GmbH
