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APPLICATION NOTE
©2017 NanoTemper Technologies, Inc. South San Francisco, CA, USA. All Rights Reserved.
Conclusion
The novel NanoTemper Technologies RED-tris-NTA
dye provides a versatile tool for efficient and site-
specific in situ labeling of His-tagged proteins. The
RED-tris-NTA labeling procedure is robust towards a
variety of different buffer conditions and components
which are o en used in storage buffers. High affinity
and selectivity of the dye for His-tags enables the
labeling of target proteins even in crude cell lysates.
Only very small amounts (picogram) of protein
are needed for the labeling and the protocol is
optimized to label only as much protein as needed
for an MST experiment. Thus, even very sensitive and
low abundant proteins can be labeled and directly
analyzed by MST without any waste of material.
Figure 4: MST traces (top) and dose-response curves (bottom) of His6-pUL53 for RED-tris-NTA (A) in the E. coli lysate and (B) the
comparison of the binding affinity of pUL50 toward His6-pUL53 measured either with purified His6-pUL53 or His6-pUL53 in
crude lysate. Measurements were performed at LED power 40% and MST power 60 %. Fnorm = normalized fluorescence
A B
