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XFD488 Phalloidin *XFD488 Same Structure to Alexa Fluor™ 488*

<strong>Fixed and stained HeLa cells.</strong><br>HeLa cells were fixed with 4% formaldehyde, permeabilized, and blocked. F-actin were stained with XFD488 phalloidin (Cat No. 23153) and nuclei labeled with Nuclear Red™ DCS1 (Cat No. 17552). Images were acquired on a Keyence BZ-X710 all-in-one fluorescence microscope.
<strong>Fixed and stained HeLa cells.</strong><br>HeLa cells were fixed with 4% formaldehyde, permeabilized, and blocked. F-actin were stained with XFD488 phalloidin (Cat No. 23153) and nuclei labeled with Nuclear Red™ DCS1 (Cat No. 17552). Images were acquired on a Keyence BZ-X710 all-in-one fluorescence microscope.
Ordering information
Price ()
Catalog Number23153
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
Physical properties
Molecular weight1300
SolventDMSO
Spectral properties
Correction Factor (260 nm)0.3
Correction Factor (280 nm)0.11
Extinction coefficient (cm -1 M -1)73000
Excitation (nm)499
Emission (nm)520
Quantum yield0.921
Storage, safety and handling
H-phraseH301, H311, H331
Hazard symbolT
Intended useResearch Use Only (RUO)
R-phraseR23, R24, R25
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200

OverviewpdfSDSpdfProtocol


Molecular weight
1300
Correction Factor (260 nm)
0.3
Correction Factor (280 nm)
0.11
Extinction coefficient (cm -1 M -1)
73000
Excitation (nm)
499
Emission (nm)
520
Quantum yield
0.921
XFD488 is manufactured by AAT Bioquest, and it has the same chemical structure of Alexa Fluor® 488 (Alexa Fluor® is the trademark of ThermoFisher). XFD488 phalloidin conjugate is chemically equivalent to Alexa Fluor® 488 phalloidin. This green fluorescent phalloidin conjugate selectively binds to F-actins. Used at nanomolar concentrations, phalloidin derivatives are convenient probes for labeling, identifying and quantitating F-actins in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments. Fluorescent phalloidin derivatives have been used as an important tool in the study of actin networks at high resolution. AAT Bioquest offers a variety of fluorescent phalloidin derivatives with different colors for multicolor imaging applications.

Example protocol


AT A GLANCE

Protocol Summary
  1. Prepare samples in microplate wells
  2. Remove liquid from samples in the plate
  3. Add XFD488 Phalloidin Conjugate solution (100 μL/well)
  4. Stain the cells at room temperature for 20 to 90 minutes
  5. Wash the cells
  6. Examine the specimen under microscope with FITC filter 
Important      Warm the vial to room temperature and centrifuge briefly before opening.

Storage and Handling Conditions
The solution should be stable for at least 6 months if store at -20 °C. Protect the fluorescent conjugates from light, and avoid freeze/thaw cycles. Note: Phalloidin is toxic, although the amount of toxin present in a vial could be lethal only to a mosquito (LD50 of phalloidin = 2 mg/kg), it should be handled with care.

PREPARATION OF WORKING SOLUTION

XFD488 Phalloidin Conjugate working solution
Add 1 µL of XFD488 Phalloidin Conjugate solution to 1 mL of PBS with 1% BSA. Note: The stock solution of phalloidin conjugate should be aliquoted and stored at -20 °C. protected from light. Note: Different cell types might be stained differently. The concentration of phalloidin conjugate working solution should be prepared accordingly.

SAMPLE EXPERIMENTAL PROTOCOL

Stain the cells
  1. Perform formaldehyde fixation. Incubate 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 XFD488 Phalloidin Conjugate working solution into the fixed cells, and stain the cells at room temperature for 20 to 90 minutes.
  5. Rinse cells gently with PBS 2 to 3 times to remove excess phalloidin conjugate before plating, sealing and imaging under microscope with FITC filter set. 

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of XFD488 Phalloidin *XFD488 Same Structure to Alexa Fluor™ 488* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM76.923 µL384.615 µL769.231 µL3.846 mL7.692 mL
5 mM15.385 µL76.923 µL153.846 µL769.231 µL1.538 mL
10 mM7.692 µL38.462 µL76.923 µL384.615 µL769.231 µL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum


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spectrum

Spectral properties

Correction Factor (260 nm)0.3
Correction Factor (280 nm)0.11
Extinction coefficient (cm -1 M -1)73000
Excitation (nm)499
Emission (nm)520
Quantum yield0.921

Citations


View all 31 citations: Citation Explorer
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Authors: Chen, Li and Tang, Ya-Li and Liu, Zhi-Hong and Pan, Ying and Jiao, Rui-Qing and Kong, Ling-Dong
Journal: European journal of pharmacology (2021): 174616
Targeted Gene Disruption in Pacific Oyster Based on CRISPR/Cas9 Ribonucleoprotein Complexes
Authors: Yu, Hong and Li, Huijuan and Li, Qi and Xu, Rui and Yue, Chenyang and Du, Shaojun
Journal: Marine Biotechnology (2019): 1--9
Enhancing the cell-biological performances of hydroxyapatite bioceramic by constructing silicate-containing grain boundary phases via sol infiltration
Authors: Xu, Yubin and Lu, Teliang and He, Fupo and Ma, Ning and Ye, Ji and ong , undefined and Wu, Tingting
Journal: ACS Biomaterials Science & Engineering (2018)
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Authors: Shi, Haishan and Ye, Xiaoling and Zhang, Jing and Ye, Ji and ong, undefined
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Authors: Xiong, Kun and Fan, Qingbo and Wu, Tingting and Shi, Haishan and Chen, Lin and Yan, Minhao
Journal: Materials Science and Engineering: C (2017)
Cell-Permeable, MMP-2 Activatable, Nickel Ferrite and His-tagged Fusion Protein Self-Assembled Fluorescent Nanoprobe for Tumor Magnetic Targeting and Imaging
Authors: Sun, Lu and Xie, Shuping and Qi, Jing and Liu, Ergang and Liu, Di and Liu, Quan and Chen, Sunhui and He, Huining and Yang, Victor C
Journal: ACS Applied Materials & Interfaces (2017)
The correlation between osteopontin adsorption and cell adhesion to mixed self-assembled monolayers of varying charges and wettability
Authors: Hao, Lijing and Li, Tianjie and Yang, Fan and Zhao, Naru and Cui, Fuzhai and Shi, Xuetao and Du, Chang and Wang, Yingjun
Journal: Biomaterials Science (2017)
Study on the Regulation of Focal Adesions and Cortical Actin by Matrix Nanotopography in 3D Environment
Authors: Han, Jingjing and Lin, Keng-hui and Chew, Lock Yue
Journal: Journal of Physics: Condensed Matter (2017)
Enhanced osteointegration of tantalum-modified titanium implants with micro/nano-topography
Authors: Shi, Junyu and Zhang, Xiaomeng and Qiao, Shichong and Ni, Jie and Mo, Jiaji and Gu, Yingxin and Lai, Hongchang
Journal: RSC Advances (2017): 46472--46479

References


View all 127 references: Citation Explorer
Improved penile histology by phalloidin stain: circular and longitudinal cavernous smooth muscles, dual-endothelium arteries, and erectile dysfunction-associated changes
Authors: Lin G, Qiu X, F and el TM, Albersen M, Wang Z, Lue TF, Lin CS.
Journal: Urology (2011): 970 e1
Phalloidin perturbs the interaction of human non-muscle myosin isoforms 2A and 2C1 with F-actin
Authors: Diensthuber RP, Muller M, Heissler SM, Taft MH, Chizhov I, Manstein DJ.
Journal: FEBS Lett (2011): 767
pH-(low)-insertion-peptide (pHLIP) translocation of membrane impermeable phalloidin toxin inhibits cancer cell proliferation
Authors: An M, Wijesinghe D, Andreev OA, Reshetnyak YK, Engelman DM.
Journal: Proc Natl Acad Sci U S A (2010): 20246
Labeling cytoskeletal F-actin with rhodamine phalloidin or fluorescein phalloidin for imaging
Authors: Chazotte B., undefined
Journal: Cold Spring Harb Protoc (2010): pdb prot4947
Protective effect of bile acid derivatives in phalloidin-induced rat liver toxicity
Authors: Herraez E, Macias RI, Vazquez-Tato J, Hierro C, Monte MJ, Marin JJ.
Journal: Toxicol Appl Pharmacol (2009): 21
Effect of Phalloidin on Filaments Polymerized from Heart Muscle Adp-Actin Monomers
Authors: Vig A, Dudas R, Kupi T, Orban J, Hild G, Lorinczy D, Nyitrai M.
Journal: J Therm Anal Calorim (2009): 721
In vitro inhibition of OATP-mediated uptake of phalloidin using bile acid derivatives
Authors: Herraez E, Macias RI, Vazquez-Tato J, Vicens M, Monte MJ, Marin JJ.
Journal: Toxicol Appl Pharmacol (2009): 13
Processing of the phalloidin proprotein by prolyl oligopeptidase from the mushroom Conocybe albipes
Authors: Luo H, Hallen-Adams HE, Walton JD.
Journal: J Biol Chem (2009): 18070
Pygmy squids and giant brains: mapping the complex cephalopod CNS by phalloidin staining of vibratome sections and whole-mount preparations
Authors: Wollesen T, Loesel R, Wanninger A.
Journal: J Neurosci Methods (2009): 63
Anti-acetylated tubulin antibody staining and phalloidin staining in the starlet sea anemone Nematostella vectensis
Authors: Genikhovich G, Technau U.
Journal: Cold Spring Harb Protoc (2009): pdb prot5283