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Neuron 98, 977–91. Excitatory and Inhibitory Neurons Utilize Different Ca2+ Sensors and Sources to Regulate Spontaneous Release. 2018

Courtney, N.A., Briguglio, J.S., Bradberry, M.M., Greer, C., Chapman, E.R.

Notes: To localize and see Doc2 in live neurons the authors generated C-terminal HaloTag® fusion proteins of Doc2 isoforms, then conjugated these fusions with JF646 HaloTag® Ligand dye. In other studies a HaloTag ® Pull-Down System to study t-SNARE binding activities on the syt1 and Doc2β constructs. (5095)

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Science 361. Imaging dynamic and selective low-complexity domain interactions that control gene transcription. 2018

Chong, S., Dugast-Darzacq, C., Liu, Z., Dong, P., Dailey, G.M., Cattoglio, C., Heckert, A., Banala, S., Lavis, L., Darzacq, X., Tjian, R.

Notes: The authors used Janelia Fluor® 549 HaloTag® and Janelia Fluor® 646 HaloTag® Ligands to stain oncogene fusion knock-ins, allowing visualization of oncogene fusion hubs in the cell nucleus and real-time tracking of individual fusion molecules. (5071)

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Nature Methods 15(6), 425–28. Single-shot super-resolution total internal reflection fluorescence microscopy. 2018

Guo, M., Chandris, P., Giannini, J.P., Trexler, A.J., Fischer, R., Chen, J., Vishwasrao, H.D., Rey-Suarez, I., Wu, Y., Wu, X., Waterman, C.M., Patterson, G.H., Upadhyaya, A., Taraska, J.W., Shroff, H.

Notes: The authors combined instant structured illumination microscopy (iSIM) with total internal reflection fluorescence microscopy (TIRFM) in an approach referred to as instant TIRF-SIM, and applied it to live samples to achieve rapid, high-contrast super-resolution imaging. Human osteosarcoma U2OS cells were transfected to monitor wild-type Ras dynamics and  imaged with a HaloTag® chimera of HRas labeled with Janelia Fluor 549. (5070)

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Methods 123, 76–88. Quantifying transcription factor binding dynamics at the single-molecule level in live cells. 2017

Presman, D.M., Ball, D.A., Paakinaho, V., Grimm, J.B., Lavis, L.D., Karpova, T.S., Hager, G.L.

Notes: The authors used single-molecule tracking (SMT) approach to follow individual protein molecules in single live cells and performed SMT experiments to learn about the binding characteristics of transcription factors (TFs) such as residence time and bound fractions. They compared JF 549 behavior on HaloTag and other tagging systems, SNAP-tag and CLIP-tag, by labeling all with JF549 dye and studying by whether the fluorophore label and/or tagging system had an effect on data obtained when using the glucocorticoid receptor (GF) as a model TF. Results showed that HaloTag is more resistant to photobleaching and enables tracking of longer binding events compared to SNAP-tag and CLIP-tag. (5069)

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Nature Methods 12, 244–50. A general method to improve fluorophores for live-cell and single-molecule microscopy. 2015

Grimm, J.B., English, B.P., Chen, J., Slaughter, J.P., Zhang., Z., Revyakin, A., Patel, R., Macklin, J.J., Normanno, D., Singer, R.H., Lionnet, T., Lavis, L.D.

Notes: Using both Janelia Fluor® 549 and 646 HaloTag® Ligands the authors compared them to existing HaloTag® ligands, known to label live cells. The new Janelia Fluor® ligands had superior brightness. (5063)

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