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Cell Navigator® Cell Plasma Membrane Staining Kit *Orange Fluorescence*
The Cell Navigator® Plasma Membrane Stain Kit provides a fast and uniform labeling of the plasma membrane without the cell-type differences exhibited by lectins. It may be used as a segmentation tool for HCS (high-content screening), as well as to stain cellular plasma membranes for standard fluorescence microscopy. The stain used in the kit survives fixation, but not permeabilization, so it is not suitable for experiments that also involve probing internal targets via antibodies.
Example protocol
AT A GLANCE
Protocol Summary
- Prepare cells in growth medium
- Incubate cells with Cellpaint™ Orange working solution at 37 ºC for 10 - 20 minutes
- Analyze the cells under fluorescence microscope at Ex/Em = 540/590 nm (TRITC filter set)
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™ Orange stock solution (500X)
Add 100 µL of DMSO (Component C) into the vial of Cellpaint™ Orange (Component A) to make 500X Cellpaint™ Orange stock solution. Note: Unused 500X Cellpaint™ Orange stock solution can be stored at ≤ -20 ºC for one month if the tubes are sealed tightly. Protect from light.PREPARATION OF WORKING SOLUTION
Add 20 µL of 500X Cellpaint™ Orange stock solution into 10 mL of Assay Buffer (Component B) and mix well to make Cellpaint™ Orange working solution. This Cellpaint™ Orange working solution is stable for at least 8 hours at room temperature. Protect from light. Note: 20 µL of 500X Cellpaint™ Orange 500X stock solution is enough for one 96-well plate.
SAMPLE EXPERIMENTAL PROTOCOL
- Add 100 µL/well (96-well plate) or 50 µL/well (384-well plate) of Cellpaint™ Orange working solution in the cell plate.
- Incubate the cells at 37°C for 10 - 20 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 Cellpaint™ Orange working solution in each well.
- Wash cells with physiological buffer (such as HHBS, PBS or buffer of your choice) for three times.
- Fix cells after staining (Optional). Fix the cells with 4% formaldehyde for 15 - 30 minutes. Wash cells with physiological buffer for three times.
- Observe the fluorescence signal in cells using a fluorescence microscope with TRITC filter set (Ex/Em = 540/590 nm).
Spectrum
Product family
| Name | Excitation (nm) | Emission (nm) |
| Cell Navigator® Cell Plasma Membrane Staining Kit *Green Fluorescence* | 497 | 505 |
| Cell Navigator® Cell Plasma Membrane Staining Kit *Red Fluorescence* | 652 | 670 |
Citations
View all 3 citations: Citation Explorer
Tissue Adhesion-Anisotropic Polyrotaxane Hydrogels Bilayered with Collagen
Authors: Hakariya, Masahiro and Arisaka, Yoshinori and Masuda, Hiroki and Yoda, Tetsuya and Tamura, Atsushi and Iwata, Takanori and Yui, Nobuhiko
Journal: Gels (2021): 168
Authors: Hakariya, Masahiro and Arisaka, Yoshinori and Masuda, Hiroki and Yoda, Tetsuya and Tamura, Atsushi and Iwata, Takanori and Yui, Nobuhiko
Journal: Gels (2021): 168
Insulin receptor based lymphocyte trafficking in the progression of type 1 diabetes
Authors: Morran, Michael P and Al-Dieri, Ali G and Nestor-Kalinoski, Andrea L and Jordan, Richard K and Gupta, Nirdesh K and McInerney, Marcia F
Journal: Journal of Biological Methods (2018): e85
Authors: Morran, Michael P and Al-Dieri, Ali G and Nestor-Kalinoski, Andrea L and Jordan, Richard K and Gupta, Nirdesh K and McInerney, Marcia F
Journal: Journal of Biological Methods (2018): e85
Inflammation in Obesity and Molecular Engineering of a Transgenic Mouse Model of Diabetes
Authors: Al-Dieri, Ali Ghalib
Journal: (2016)
Authors: Al-Dieri, Ali Ghalib
Journal: (2016)
References
View all 6 references: Citation Explorer
Activation of the neurokinin 3 receptor promotes filopodia growth and sprouting in rat embryonic hypothalamic cells
Authors: Flynn FW, Kinney-Lang E, Hoekstra C, Pratt DL, Thakar A.
Journal: Dev Neurobiol (2015): 12
Authors: Flynn FW, Kinney-Lang E, Hoekstra C, Pratt DL, Thakar A.
Journal: Dev Neurobiol (2015): 12
Probing endocytosis from the enterocyte brush border using fluorescent lipophilic dyes: lipid sorting at the apical cell surface
Authors: Danielsen EM., undefined
Journal: Histochem Cell Biol (2015): 545
Authors: Danielsen EM., undefined
Journal: Histochem Cell Biol (2015): 545
Imaging fenestrations in liver sinusoidal endothelial cells by optical localization microscopy
Authors: Monkemoller V, Schuttpelz M, McCourt P, Sorensen K, Smedsrod B, Huser T.
Journal: Phys Chem Chem Phys (2014): 12576
Authors: Monkemoller V, Schuttpelz M, McCourt P, Sorensen K, Smedsrod B, Huser T.
Journal: Phys Chem Chem Phys (2014): 12576
Effects of corticotrophin releasing hormone (CRH) on cell viability and differentiation in the human BeWo choriocarcinoma cell line: a potential syncytialisation inducer distinct from cyclic adenosine monophosphate (cAMP)
Authors: Chen Y, Allars M, Pan X, Maiti K, Angeli G, Smith R, Nicholson RC.
Journal: Reprod Biol Endocrinol (2013): 30
Authors: Chen Y, Allars M, Pan X, Maiti K, Angeli G, Smith R, Nicholson RC.
Journal: Reprod Biol Endocrinol (2013): 30
Evaluation of high-throughput screening for in vitro micronucleus test using fluorescence-based cell imaging
Authors: Shibai-Ogata A, Kakinuma C, Hioki T, Kasahara T.
Journal: Mutagenesis (2011): 709
Authors: Shibai-Ogata A, Kakinuma C, Hioki T, Kasahara T.
Journal: Mutagenesis (2011): 709
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