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Cell Navigator® Cell Plasma Membrane Staining Kit *Green 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 green fluorescent 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
- Prepare and add Cellpaint™ Green working solution to cells
- Incubate at 37°C for 5 to 20 minutes
- Read fluorescence intensity with FITC filter set
Note This protocol only provides a guideline and should be modified according to your specific needs.
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.
Note 20 uL of Cellpaint™ Green 500X stock solution is enough for one 96-well plate. Unused Cellpaint™ Green 500X stock solution can be aliquoted and stored at ≤ -20°C for one month if the tubes are sealed tightly. Protect from light and avoid repeated freeze-thaw cycles.
Cellpaint™ Green stock solution (500X)
Add 100 uL of DMSO (Component C) into the vial of Cellpaint™ Green (Component A) to make 500X stock solution. Note 20 uL of Cellpaint™ Green 500X stock solution is enough for one 96-well plate. Unused Cellpaint™ Green 500X stock solution can be aliquoted and stored at ≤ -20°C for one month if the tubes are sealed tightly. Protect from light and avoid repeated freeze-thaw cycles.
PREPARATION OF WORKING SOLUTION
Cellpaint™ Green working solution (1X)
Add 20 uL of 500X stock solution into 10 mL of Assay Buffer (Component B), and mix well. Note We recommend making the working solution fresh before use.
SAMPLE EXPERIMENTAL PROTOCOL
- Add 100 uL/well (96-well plate) or 50 uL/well (384-well plate) of Cellpaint™ Green working solution in the cell plate. Incubate the cells at 37°C for 5-20 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 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.
- Optional: Fix cells after staining. 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 fluorescence microscope with a FITC filter set.
Spectrum
Product family
| Name | Excitation (nm) | Emission (nm) |
| Cell Navigator® Cell Plasma Membrane Staining Kit *Orange Fluorescence* | 555 | 573 |
| Cell Navigator® Cell Plasma Membrane Staining Kit *Red Fluorescence* | 652 | 670 |
Citations
View all 33 citations: Citation Explorer
Matrix Rigidity-Dependent Regulation of Ca(2+) at Plasma Membrane Microdomains by FAK Visualized by Fluorescence Resonance Energy Transfer
Authors: Kim, T. J., Lei, L., Seong, J., Suh, J. S., Jang, Y. K., Jung, S. H., Sun, J., Kim, D. H., Wang, Y.
Journal: Adv Sci (Weinh) (2019): 1801290
Authors: Kim, T. J., Lei, L., Seong, J., Suh, J. S., Jang, Y. K., Jung, S. H., Sun, J., Kim, D. H., Wang, Y.
Journal: Adv Sci (Weinh) (2019): 1801290
Revealing the Raft Domain Organization in the Plasma Membrane by Single-Molecule Imaging of Fluorescent Ganglioside Analogs
Authors: Suzuki, K. G. N., Ando, H., Komura, N., Konishi, M., Imamura, A., Ishida, H., Kiso, M., Fujiwara, T. K., Kusumi, A.
Journal: Methods Enzymol (2018): 267-282
Authors: Suzuki, K. G. N., Ando, H., Komura, N., Konishi, M., Imamura, A., Ishida, H., Kiso, M., Fujiwara, T. K., Kusumi, A.
Journal: Methods Enzymol (2018): 267-282
A single fluorescent probe enables clearly discriminating and simultaneously imaging liquid-ordered and liquid-disordered microdomains in plasma membrane of living cells
Authors: Tian, M., Liu, Y., Sun, Y., Zhang, R., Feng, R., Zhang, G., Guo, L., Li, X., Yu, X., Sun, J. Z., He, X.
Journal: Biomaterials (2017): 46-56
Authors: Tian, M., Liu, Y., Sun, Y., Zhang, R., Feng, R., Zhang, G., Guo, L., Li, X., Yu, X., Sun, J. Z., He, X.
Journal: Biomaterials (2017): 46-56
Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
Authors: Ji, C., Lou, X.
Journal: J Vis Exp (2016): se name="22682.enl" path="C:\Users\aatbi\Drop
Authors: Ji, C., Lou, X.
Journal: J Vis Exp (2016): se name="22682.enl" path="C:\Users\aatbi\Drop
Determination of Dynamics of Plant Plasma Membrane Proteins with Fluorescence Recovery and Raster Image Correlation Spectroscopy
Authors: Lankova, M., Humpolickova, J., Vosolsobe, S., Cit, Z., Lacek, J., Covan, M., Covanova, M., Hof, M., Petrasek, J.
Journal: Microsc Microanal (2016): 290-9
Authors: Lankova, M., Humpolickova, J., Vosolsobe, S., Cit, Z., Lacek, J., Covan, M., Covanova, M., Hof, M., Petrasek, J.
Journal: Microsc Microanal (2016): 290-9
References
View all 6 references: Citation Explorer
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
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
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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