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Cell Navigator® Cell Plasma Membrane Staining Kit *Red 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
  1. Prepare cells in growth medium
  2. Incubate cells with Cellpaint™ Deep Red working solution at 37°C for 10 - 20 minutes
  3. Analyze the cells under fluorescence microscope at Ex/Em = 640/660 nm (Cy5 filter set) 
Important      Thaw all the kit components at room temperature before starting the experiment.

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™ Deep Red stock solution (500X)
Add 100 µL of DMSO (Component C) into the vial of Cellpaint™ Deep Red (Component A) to make 500X Cellpaint™ Deep Red stock solution. Note: Protect from light. For storage, seal tubes tightly.

PREPARATION OF WORKING SOLUTION

Add 20 µL of 500X Cellpaint™ Deep Red stock solution into 10 mL of Assay Buffer (Component B), and mix well to make Cellpaint™ Deep Red working solution. This Cellpaint™ Deep Red working solution is stable for at least 8 hours at room temperature. Protect from light. Note: 20 µL of 500X Cellpaint™ Deep Red 500X stock solution is enough for one 96-well plate.

SAMPLE EXPERIMENTAL PROTOCOL

  1. Add 100 µL/well (96-well plate) or 50 µL/well (384-well plate) of Cellpaint™ Deep Red working solution in the cell plate.
  2. 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.
  3. Remove Cellpaint™ Deep Red working solution in each well.
  4. Wash cells with physiological buffer (such as HHBS, PBS or buffer of your choice) for three times.
  5. Fix cells after staining (Optional). Fix the cells with 4% formaldehyde for 15 - 30 minutes. Wash cells with physiological buffer for three times.
  6. Observe the fluorescence signal in cells using a fluorescence microscope with Cy5 filter set (Ex/Em =640/660 nm). 

Spectrum

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
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
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
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
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
Page updated on December 1, 2024

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Catalog Number22681
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Spectral properties

Excitation (nm)

652

Emission (nm)

670

Storage, safety and handling

Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200

Platform

Fluorescence microscope

ExcitationCy5 filter
EmissionCy5 filter
Recommended plateBlack wall, clear bottom

Components