logo
AAT Bioquest

Cell Navigator® F-Actin Labeling Kit *Green Fluorescence*

Fluorescence image of HeLa cells fixed with 4% formaldehyde then stained with Cell Navigator® F-Actin Labeling Kit *Green Fluorescence* in a Costar black 96-well plate. Cell were labeled with iFluor® 488-Phalloidin (Cat#22261, Green) and nuclei stain DAPI (Cat#17507, Blue), respectively. Cell endoplasmic reticulum (ER) was stained with ER Red™ (Cat#22636, Red) before fixation.
Fluorescence image of HeLa cells fixed with 4% formaldehyde then stained with Cell Navigator® F-Actin Labeling Kit *Green Fluorescence* in a Costar black 96-well plate. Cell were labeled with iFluor® 488-Phalloidin (Cat#22261, Green) and nuclei stain DAPI (Cat#17507, Blue), respectively. Cell endoplasmic reticulum (ER) was stained with ER Red™ (Cat#22636, Red) before fixation.
Fluorescence image of HeLa cells fixed with 4% formaldehyde then stained with Cell Navigator® F-Actin Labeling Kit *Green Fluorescence* in a Costar black 96-well plate. Cell were labeled with iFluor® 488-Phalloidin (Cat#22261, Green) and nuclei stain DAPI (Cat#17507, Blue), respectively. Cell endoplasmic reticulum (ER) was stained with ER Red™ (Cat#22636, Red) before fixation.
Ordering information
Price
Catalog Number
Unit Size
Quantity
Add to cart
Additional ordering information
Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
InternationalSee distributors
Bulk requestInquire
Custom sizeInquire
ShippingStandard overnight for United States, inquire for international
Request quotation
Spectral properties
Correction Factor (260 nm)0.21
Correction Factor (280 nm)0.11
Extinction coefficient (cm -1 M -1)750001
Excitation (nm)491
Emission (nm)516
Quantum yield0.91
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Related products
Cell Navigator® Live Cell RNA Imaging Kit *Green Fluorescence*
Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit *Green Fluorescence*
Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit *Red Fluorescence*
Cell Navigator® Lysosome Staining Kit *Green Fluorescence with 405 nm Excitation*
Cell Navigator® Lysosome Staining Kit *NIR Fluorescence*
Cell Navigator® Lysosome Staining Kit *Blue Fluorescence*
Cell Navigator® Lysosome Staining Kit *Green Fluorescence*
Cell Navigator® Lysosome Staining Kit *Orange Fluorescence*
Cell Navigator® Lysosome Staining Kit *Red Fluorescence*
Cell Navigator® Lysosome Staining Kit *Deep Red Fluorescence*
Cell Navigator® Mitochondrion Staining Kit *Blue Fluorescence*
Cell Navigator® Mitochondrion Staining Kit *Green Fluorescence*
Cell Navigator® Mitochondrion Staining Kit *Orange Fluorescence*
Cell Navigator® Mitochondrion Staining Kit *Red Fluorescence*
Cell Navigator® Mitochondrion Staining Kit *Deep Red Fluorescence*
Cell Navigator® Mitochondrion Staining Kit *Orange Fluorescence with 405 nm Excitation*
Cell Navigator® Cell Plasma Membrane Staining Kit *Orange Fluorescence*
Cell Navigator® Cell Plasma Membrane Staining Kit *Red Fluorescence*
Cell Navigator® Mitochondrion Staining Kit *NIR Fluorescence*
Cell Navigator® Fluorimetric Lipid Droplet Assay Kit *Green Fluorescence*
Cell Navigator® Fluorimetric Lipid Droplet Assay Kit *Red Fluorescence*
Cell Navigator® NBD Ceramide Golgi Staining Kit *Green Fluorescence*
Cell Navigator® Live Cell Tubulin Staining Kit
Cell Navigator® Cell Plasma Membrane Staining Kit *Green Fluorescence*
Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit *Blue Fluorescence*
Cell Navigator® TMR Ceramide Golgi Staining Kit *Red Fluorescence*
Cell Navigator® CDy6 Mitosis Imaging Kit
Show More (17)

OverviewpdfSDSpdfProtocol


Correction Factor (260 nm)
0.21
Correction Factor (280 nm)
0.11
Extinction coefficient (cm -1 M -1)
750001
Excitation (nm)
491
Emission (nm)
516
Quantum yield
0.91
Our Cell Navigator® fluorescence imaging kits are a set of fluorescence imaging tools for labeling sub-cellular organelles such as membranes, lysosomes, mitochondria and nuclei etc. The selective labeling of live cell compartments provides a powerful method for studying cellular events in a spatial and temporal context. This particular kit is designed to label F-actins of fixed cells in green fluorescence. The kit uses a green fluorescent phalloidin conjugate that is selectively bound to F-actins. This green fluorescent phalloidin conjugate is a high-affinity probe for F-actins with much higher photostability than the fluorescein-phalloidin conjugates. Used at nanomolar concentrations, phallotoxins are convenient probes for labeling, identifying and quantitating F-actins in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments. The labeling protocol is robust, requiring minimal hands-on time. The kit provides all the essential components with an optimized staining protocol.

Platform


Fluorescence microscope

ExcitationFITC filter
EmissionFITC filter
Recommended plateBlack wall/clear bottom

Components


Example protocol


AT A GLANCE

Protocol summary

  1. Prepare samples (microplate wells)
  2. Remove the liquid from the plate
  3. Add 100 µL/well of iFluor™ 488-Phalloidin working solution
  4. Stain the cells at RT for 15 to 60 minutes
  5. Wash the cells
  6. Examine the specimen under fluorescence microscope at Ex/Em = 490/520 nm (FITC filter set)

Important notes
Thaw all the components at room temperature before starting the experiment.

PREPARATION OF WORKING SOLUTION

Add 10 μL of iFluor™ 488-Phalloidin (Component A) to 10 mL of Labeling Buffer (Component B) to make 1X iFluor™ 488-Phalloidin working solution. Protect from light. Note: Different cell types might be stained differently. The concentration of iFluor™ 488-Phalloidin working solution should be prepared accordingly.

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

SAMPLE EXPERIMENTAL PROTOCOL

  1. Perform formaldehyde fixation. Incubate the cells with 3.0% – 4.0% formaldehyde in PBS at room temperature for 10 – 30 minutes. Note: Avoid any methanol containing fixatives since methanol can disrupt actin during the fixation process. The preferred fixative is methanol-free formaldehyde.

  2. Rinse the fixed cells 2 – 3 times in PBS.

  3. Optional: Add 0.1% Triton X-100 in PBS into fixed cells for 3 to 5 minutes to increase permeability. Rinse the cells 2 – 3 times in PBS.

  4. Add 100 µL/well (96-well plate) of iFluor™ 488-Phalloidin working solution into the fixed cells.

  5. Stain the cells at room temperature for 15 to 60 minutes.

  6. Rinse cells gently with PBS 2 to 3 times to remove excess dye before plate sealing.

  7. Image cells using a fluorescence microscope with FITC filter set (Ex/Em = 490/520 nm).

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Correction Factor (260 nm)0.21
Correction Factor (280 nm)0.11
Extinction coefficient (cm -1 M -1)750001
Excitation (nm)491
Emission (nm)516
Quantum yield0.91

Product Family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
Cell Navigator® F-Actin Labeling Kit *Blue Fluorescence*3454502000010.9510.830.23
Cell Navigator® F-Actin Labeling Kit *Orange Fluorescence*54155710000010.6710.250.15
Cell Navigator® F-Actin Labeling Kit *Red Fluorescence*58760320000010.5310.050.04

Images


Citations


View all 40 citations: Citation Explorer
TRPA1 and TPRV1 Ion Channels Are Required for Contact Lens-Induced Corneal Parainflammation and Can Modulate Levels of Resident Corneal Immune Cells
Authors: Datta, Ananya and Lee, Ji Hyun and Flandrin, Orneika and Horneman, Hart and Lee, Justin and Metruccio, Matteo ME and Bautista, Diana and Evans, David J and Fleiszig, Suzanne MJ
Journal: Investigative Ophthalmology \& Visual Science (2023): 21--21
Modification of adipose mesenchymal stem cells-derived small extracellular vesicles with fibrin-targeting peptide CREKA for enhanced bone repair
Authors: Wu, Qi and Fu, Xiaoling and Li, Xian and Li, Jing and Han, Weiju and Wang, Yingjun
Journal: Bioactive Materials (2023): 208--220
Microfluidic Chip-Based Modeling of Three-Dimensional Intestine--Vessel--Liver Interactions in Fluorotelomer Alcohol Biotransformation
Authors: Xu, Ning and Lin, Haifeng and Lin, Jin-Ming and Cheng, Jie and Wang, Peilong and Lin, Ling
Journal: Analytical Chemistry (2023)
Digital light processing-bioprinted poly-NAGA-GelMA-based hydrogel lenticule for precise refractive errors correction
Authors: Jia, Shuo and Yang, Jirong and Lau, Aaron Dzi-Shing and Chen, Fushun and Bu, Yashan and Cai, Erlong and Wang, Huogang and Chieng, Herng-Ee and Sun, Tianhao and Zhiyong, Zhou and others,
Journal: Biofabrication (2023)
Honeycomb Biosilica in Sponges: From Understanding Principles of Unique Hierarchical Organization to Assessing Biomimetic Potential
Authors: Voronkina, Alona and Romanczuk-Ruszuk, Eliza and Przekop, Robert E and Lipowicz, Pawel and Gabriel, Ewa and Heimler, Korbinian and Rogoll, Anika and Vogt, Carla and Frydrych, Milosz and Wienclaw, Pawel and others,
Journal: Biomimetics (2023): 234
Adjusting physicochemical and cytological properties of biphasic calcium phosphate by magnesium substitution: An in vitro study
Authors: Lu, Teliang and Miao, Yali and Yuan, Xinyuan and Zhang, Yu and Ye, Jiandong
Journal: Ceramics International (2023)
CircRPAP2 regulates the alternative splicing of PTK2 by binding to SRSF1 in breast cancer
Authors: Yu, Yunhe and Fang, Lin
Journal: Cell death discovery (2022): 1--12
Cholesterol inhibits autophagy in RANKL-induced osteoclast differentiation through activating the PI3K/AKT/mTOR signaling pathway
Authors: Jiang, Chunyan and Wang, Yan and Zhang, Mengqi and Xu, Jin
Journal: Molecular Biology Reports (2022): 9217--9229
A targetable pathway in neutrophils mitigates both arterial and venous thrombosis
Authors: Nayak, Lalitha and Sweet, David R and Thomas, Asha and Lapping, Stephanie D and Kalikasingh, Kenneth and Madera, Annmarie and Vinayachandran, Vinesh and Padmanabhan, Roshan and Vasudevan, Neelakantan T and Myers, Jay T and others,
Journal: Science Translational Medicine (2022): eabj7465

References


View all 42 references: Citation Explorer
Velocity distributions of single F-actin trajectories from a fluorescence image series using trajectory reconstruction and optical flow mapping
Authors: von Wegner F, Ober T, Weber C, Schurmann S, Winter R, Friedrich O, Fink RH, Vogel M.
Journal: J Biomed Opt (2008): 54018
Visualization of F-actin and G-actin equilibrium using fluorescence resonance energy transfer (FRET) in cultured cells and neurons in slices
Authors: Okamoto K, Hayashi Y.
Journal: Nat Protoc (2006): 911
The effect of F-actin on the relay helix position of myosin II, as revealed by tryptophan fluorescence, and its implications for mechanochemical coupling
Authors: Conibear PB, Malnasi-Csizmadia A, Bagshaw CR.
Journal: Biochemistry (2004): 15404
Analysis of models of F-actin using fluorescence resonance energy transfer spectroscopy
Authors: Moens PD, dos Remedios CG.
Journal: Results Probl Cell Differ (2001): 59
Fluorescence studies of the carboxyl-terminal domain of smooth muscle calponin effects of F-actin and salts
Authors: Bartegi A, Roustan C, Kassab R, Fattoum A.
Journal: Eur J Biochem (1999): 335
Microquantification of cellular and in vitro F-actin by rhodamine phalloidin fluorescence enhancement
Authors: Katanaev VL, Wymann MP.
Journal: Anal Biochem (1998): 185
A conformational change in F-actin when myosin binds: fluorescence resonance energy transfer detects an increase in the radial coordinate of Cys-374
Authors: Moens PD, dos Remedios CG.
Journal: Biochemistry (1997): 7353
Interhead distances in myosin attached to F-actin estimated by fluorescence energy transfer spectroscopy
Authors: Ishiwata S, Miki M, Shin I, Funatsu T, Yasuda K, dos Remedios CG.
Journal: Biophys J (1997): 895
Myosin-induced changes in F-actin: fluorescence probing of subdomain 2 by dansyl ethylenediamine attached to Gln-41
Authors: Kim E, Miller CJ, Motoki M, Seguro K, Muhlrad A, Reisler E.
Journal: Biophys J (1996): 1439
Changes in the distribution of F-actin in the fission yeast Schizosaccharomyces pombe by arresting growth in distilled water: correlative studies with fluorescence and electron microscopy
Authors: Kanbe T, Akashi T, Tanaka K.
Journal: J Electron Microsc (Tokyo) (1994): 20