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CellPaint™ TSP membrane stain
TSP is a styrylpyridine-based fluorescent membrane probe suitable for imaging plasma membranes in living cells and tissues. It was reported by Guo et al in 2016 (Analyst, 2016, 141, 3228). The probe is a molecular rotor that has fluorescence sharply enhanced in viscous media. Its fluorescence is also microenvironment-sensitive, enables the turn-on imaging of plasma membranes with a high signal-to-noise ratio. Guo et al has demonstrated that TSP has high photostability, low cytotoxicity and excellent biocompatibility. It can also be used in 2 photon imaging.
Example protocol
AT A GLANCE
- Prepare cells in growth medium
- Incubate cells with Cellpaint™ TSP membrane stain
- Analyze under fluorescence microscope with Cy3/TRITC filter set
Chemical and Physical Properties
Molecular Weight: 1016.27Solvent: dimethyl sulfoxide (DMSO)
Spectral Properties: Excitation = 490 nm; Emission = 600 nm
CELL PREPARATION
For guidelines on cell sample preparation, please visit https://www.aatbio.com/resources/guides/cell-sample-preparation.html
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
Cellpaint™ TSP membrane stain stock solution (500X)
- Thaw the vial of CellPaint™ TSP membrane stain at room temperature before use.
- Add 100 µL of DMSO into the vial of Cellpaint™ TSP membrane stain to make 500X stock solution.
Note Unused Cellpaint™ TSP membrane stain stock solution can be aliquoted and stored at ≤ -20 °C for a couple of months if the tubes are sealed tightly. Protect from light and avoid repeated freeze-thaw cycles.
PREPARATION OF WORKING SOLUTION
Cellpaint™ TSP membrane stain working solution (10X)
- Add 20 µL of 500X stock solution into 1 mL of cell culture medium, and mix well.
Note We recommend making the working solution fresh before use.
Note 20 µL of 500X Cellpaint™ TSP membrane stain stock solution is enough for one 96-well plate.
Note We recommend using a cell culture medium to make a working solution for better resolution images.
SAMPLE EXPERIMENTAL PROTOCOL
Stain Cells
- Add 10 µL/well (96-well plate/100 µL volume) or 5 µL/well (384-well plate-50 µL volume) of 10X Cellpaint™ TSP membrane stain working solution in the cell plate. Incubate the cells at 37 °C for 30-60 minutes, protected from light.
Note The optimal concentration of the cell membrane probe varies depending on the specific application. The staining conditions may be modified according to the particular cell type and the permeability of the cells or tissues to the probe. - Remove working solution in each well. Wash cells with physiological buffer (such as HHBS, DPBS or buffer of your choice) for three times and replace with HHBS.
- Observe the fluorescence signal in cells using fluorescence microscope with a Cy3/TRITC filter set.
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of CellPaint™ TSP membrane stain to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
| 0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
| 1 mM | 98.399 µL | 491.995 µL | 983.99 µL | 4.92 mL | 9.84 mL |
| 5 mM | 19.68 µL | 98.399 µL | 196.798 µL | 983.99 µL | 1.968 mL |
| 10 mM | 9.84 µL | 49.2 µL | 98.399 µL | 491.995 µL | 983.99 µL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
| Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum
Citations
View all 31 citations: Citation Explorer
Membrane trafficking and exocytosis are upregulated in port wine stain blood vessels
Authors: Yin, R., Rice, S. J., Wang, J., Gao, L., Tsai, J., Anvari, R. T., Zhou, F., Liu, X., Wang, G., Tang, Y., Mihm, M. C., Jr., Belani, C. P., Chen, D. B., Nelson, J. S., Tan, W.
Journal: Histol Histopathol (2019): 479-490
Authors: Yin, R., Rice, S. J., Wang, J., Gao, L., Tsai, J., Anvari, R. T., Zhou, F., Liu, X., Wang, G., Tang, Y., Mihm, M. C., Jr., Belani, C. P., Chen, D. B., Nelson, J. S., Tan, W.
Journal: Histol Histopathol (2019): 479-490
Cu(2+)-Directed Liposome Membrane Fusion, Positive-Stain Electron Microscopy, and Oxidation
Authors: Liu, Y., Liu, J.
Journal: Langmuir (2018): 7545-7553
Authors: Liu, Y., Liu, J.
Journal: Langmuir (2018): 7545-7553
Extraction of DNA from Sperm Cells in Mixed Stain by Nylon Membrane Bushing Separation Technique
Authors: Ma, J., Tong, Q., Gao, L. B., Zhu, C., Jiang, Z. Q.
Journal: Fa Yi Xue Za Zhi (2018): 417-419
Authors: Ma, J., Tong, Q., Gao, L. B., Zhu, C., Jiang, Z. Q.
Journal: Fa Yi Xue Za Zhi (2018): 417-419
Gram's Stain Does Not Cross the Bacterial Cytoplasmic Membrane
Authors: Wilhelm, M. J., Sheffield, J. B., Sharifian Gh, M., Wu, Y., Spahr, C., Gonella, G., Xu, B., Dai, H. L.
Journal: ACS Chem Biol (2015): 1711-7
Authors: Wilhelm, M. J., Sheffield, J. B., Sharifian Gh, M., Wu, Y., Spahr, C., Gonella, G., Xu, B., Dai, H. L.
Journal: ACS Chem Biol (2015): 1711-7
The use of SMALPs as a novel membrane protein scaffold for structure study by negative stain electron microscopy
Authors: Postis, V., Rawson, S., Mitchell, J. K., Lee, S. C., Parslow, R. A., Dafforn, T. R., Baldwin, S. A., Muench, S. P.
Journal: Biochim Biophys Acta (2015): 496-501
Authors: Postis, V., Rawson, S., Mitchell, J. K., Lee, S. C., Parslow, R. A., Dafforn, T. R., Baldwin, S. A., Muench, S. P.
Journal: Biochim Biophys Acta (2015): 496-501
Page updated on November 21, 2024
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