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AAT Bioquest

Phalloidin Conjugates

XFD488 Same Structure to Alexa Fluor™ 488
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. 

Spectrum

Product family

Citations

View all 31 citations: Citation Explorer
Atractylodin inhibits fructose-induced human podocyte hypermotility via anti-oxidant to down-regulate TRPC6/p-CaMK4 signaling
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
Dicalcin suppresses in vitro trophoblast attachment in human cell lines
Authors: Saito, Ryohei and Satoh, Hiromasa and Aoba, Kayo and Hirasawa, Hajime and Miwa, Naofumi
Journal: Biochemical and Biophysical Research Communications (2021): 206--213
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)
Enhanced Osteogenesis of Injectable Calcium Phosphate Bone Cement Mediated by Loading Chondroitin Sulfate
Authors: Shi, Haishan and Ye, Xiaoling and Zhang, Jing and Ye, Ji and ong, undefined
Journal: ACS Biomaterials Science & Engineering (2018)

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
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
Page updated on October 9, 2024

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Physical properties

Molecular weight

~1300

Solvent

DMSO

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 yield

0.921

Storage, safety and handling

H-phraseH301, H311, H331
Hazard symbolT
Intended useResearch Use Only (RUO)
R-phraseR23, R24, R25

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12352200
<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&trade; 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&trade; 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&trade; DCS1 (Cat No. 17552). Images were acquired on a Keyence BZ-X710 all-in-one fluorescence microscope.