Synthesis of practical red fluorescent probe for cytoplasmic calcium ions with greatly improved cell-membrane permeability

In this data article, we described the detailed synthetic procedure and the experimental data for the synthesis of a red-fluorescent probe for calcium ions (Ca2+) with improved water solubility. This Ca2+ red-fluorescent probe CaTM-3 AM could be applied to fluorescence imaging of physiological Ca2+ concentration changes in not only live cells, but also brain slices, with high cell-membrane permeability leading to bright fluorescence in biosamples. The data provided herein are in association with the research article “The Development of Practical Red Fluorescent Probe for Cytoplasmic Calcium Ions with Greatly Improved Cell-membrane Permeability” in Cell Calcium (Hirabayashi et al., 2016) [1].


Subject area
Chemistry More specific subject area

Synthesis of fluorescent probes
Type of data Synthetic schemes, experimental synthesis protocols, NMR and MS spectra, HPLC chromatogram How data was acquired Starting compounds were either purchased or synthesized using already published synthetic protocols Experimental features Compounds were synthesized and their structures were identified by NMR and mass spectrometry Data source location

Tokyo, Japan
Data accessibility Data are provided with this article

Value of the data
The data allows to reproduce the experiments described in the research article Ref. [1]. The synthesized compound, CaTM-3 AM, could be used to fluorescence imaging of the physiological Ca 2 þ concentration change.
The data provides an opportunity to synthesize red fluorescent probes based on TokyoMagenta (TM) derivatives.

Data
NMR spectra were recorded on a JEOL JNM-LA300 instrument at 300 MHz for 1 H NMR and at 75 MHz for 13 C NMR or JEOL JNM-LA400 instrument at 100 MHz for 13 C NMR. Mass spectra (ESI þ ) were measured with a JEOL JMS-T100LC AccuTOF for ESI. HPLC analyses were performed on an Inertsil ODS-3 (4.6 Â 250 mm) column (GL Sciences Inc.) using a HPLC system composed of a pump (PU-2080, JASCO) and a detector (MD-2018 or FP-2025, JASCO).

Synthetic materials and instrumentation
General chemicals were of the best grade available, supplied by Tokyo Chemical Industries, Wako Pure Chemical, Aldrich Chemical Co., Kanto Chemical Co., Inc., Toronto Research Chemicals Inc., Watanabe Chemical Industries, Applied Biosystems, Dojindo and Invitrogen Corp., and were used without further purification. All solvents were used after appropriate distillation or purification. Preparative HPLC were performed on an Inertsil ODS-3 (10 Â 250 mm) column (GL Sciences Inc.) using a HPLC system composed of a pump (PU-2080, JASCO) and a detector (MD-2015 or FP-2025, JASCO) or a HPLC system composed of a pump (PU-2086, JASCO) and a detector (MD-2018, JASCO).

Synthesis of compound 1
This compound was synthesized according to Ref. [2].