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TECHNICAL NOTE Tycho is a trademark of NanoTemper Technologies GmbH, Munich, Germany. NanoTemper is a registered trademark of NanoTemper Technologies GmbH, Munich, Germany. ©2017 NanoTemper Technologies, Inc. South San Francisco, CA, USA. All Rights Reserved. TE-TY-003-01 nanotempertech.com References 1. Shen, X., Ranallo, R., Choi, E. & Wu, C. Involvement of actin-related proteins in ATP-dependent chromatin remodeling. Mol Cell 12, 147-55 (2003). 2. Szerlong, H. et al. The HSA domain binds nuclear actin-related proteins to regulate chromatin-remodeling ATPases. Nat Struct Mol Biol 15, 469-76 (2008). 3. Fenn et al., Structural biochemistry of nuclear actin-related proteins 4 and 8 reveals their interaction with actin. EMBO J. Jun 1;30(11):2153-66 (2011) 4. Gerhold, C.B. et al. Structure of Actin-related protein 8 and its contribution to nucleosome binding. Nucleic Acids Res 40, 11036-46 (2012). 30 40 50 60 70 80 90 100 0.84 0.86 0.88 0.90 0.92 0.94 0.96 0.98 Temperature (°C) Ratio 350 nm / 330 nm HSA complex HSA complex + 1 mM ATP Initial ratio T i (°C) Profile similarity (%) Ratio Sample Conclusions Tycho NT.6 is a powerful technology for researchers to perform effortless, accurate and robust quality checks on the folding state and functionality of their protein samples during purification. Generating results in three minutes, researchers can make faster and more educated decisions in their purification workflow. Binding interactions with the respective ligands can be quickly tested before investing time, effort and expenses in other methodologies. Figure 2: Functional validation of purified HSAX complex by thermal shi analysis. HSAX in presence and absence of 1 mM ATP was monitored using the Tycho NT.6. In the presence of 1 mM ATP, the unfolding transitions of the HSAX complex are different than for the complex alone. A summary of the results is shown in the table. Initial ratio is the initial value (at 35 °C) of the ratio of measurements collected at 350 and 330 nm. ∆ Ratio: Difference between the ratio at the beginning and at the end of the thermal profile. T i : Inflection temperature of the unfolding transition in the 350 nm / 330 nm ratio signal. Profile similarity, is an index which quantifies the sameness of the unfolding profiles of two or more samples.