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Cell Navigator™ Cell Plasma Membrane Staining Kit *Orange Fluorescence*

Fluorescence images of HL-60 cells stained with Cell Navigator™ Cell Plasma Membrane Staining Kit *Orange Fluorescence* in a 96-well black wall/clear bottom plate. The cells were imaged using a fluorescence microscope equipped with a TRITC filter.
Fluorescence images of HL-60 cells stained with Cell Navigator™ Cell Plasma Membrane Staining Kit *Orange Fluorescence* in a 96-well black wall/clear bottom plate. The cells were imaged using a fluorescence microscope equipped with a TRITC filter.
HeLa cells were labeled with Cell Navigator™ Cell Plasma Membrane Staining Kit *Orange Fluorescence* (Cat No. 22680), and nuclei were labeled with Hoechst 33342 (Cat No. 17530). Labeled cells were imaged on the Keyence BZ-X710 All-In-One Fluorescence Microscope.
Ordering information
Price ()
Catalog Number22680
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Spectral properties
Excitation (nm)555
Emission (nm)573
Storage, safety and handling
Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200

OverviewpdfSDSpdfProtocol


Excitation (nm)
555
Emission (nm)
573
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.

Platform


Fluorescence microscope

ExcitationTRITC filter
EmissionTRITC filter
Recommended plateBlack wall/clear bottom

Components


Component A: Cellpaint™ Orange1 vial
Component B: Assay Buffer1 bottle (50 mL)
Component C: DMSO1 vial (200 µL)

Example protocol


AT A GLANCE

Protocol Summary
  1. Prepare cells in growth medium
  2. Incubate cells with Cellpaint™ Orange working solution at 37 ºC for 10 - 20 minutes
  3. Analyze the cells under fluorescence microscope at Ex/Em = 540/590 nm (TRITC 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™ 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

  1. Add 100 µL/well (96-well plate) or 50 µL/well (384-well plate) of Cellpaint™ Orange 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™ Orange 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 TRITC filter set (Ex/Em = 540/590 nm). 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)555
Emission (nm)573

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
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
Inflammation in Obesity and Molecular Engineering of a Transgenic Mouse Model of Diabetes
Authors: Al-Dieri, Ali Ghalib
Journal: (2016)

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
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
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
A rapid, inexpensive high throughput screen method for neurite outgrowth
Authors: Yeyeodu ST, Witherspoon SM, Gilyazova N, Ibeanu GC.
Journal: Curr Chem Genomics (2010): 74