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Phalloidin (FITC)

Actin ligand
 
ALX-350-268-MC01 0.1 mg 235.00 USD
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For detection of microfilaments in the light microscope. Mixture of 4 isomers with lower affinity to actin than phalloidin (Prod. No. ALX-350-265).

Product Specification

Alternative Name:Fluoresceinyl-aminomethyldithiolano-phalloidin
 
Formula:C58H62N10O14S4
 
MW:1251.4
 
Source:Semisynthetic from aminomethyldithiolano-phalloidin.
 
Purity:≥90% (HPLC)
 
Appearance:Amorphous solid.
 
Solubility:Soluble in DMSO, dimethyl formamide, methanol or water (warm).
 
Shipping:Ambient
 
Long Term Storage:-20°C
 
Use/Stability:Prepare fresh solutions for each application.
 
Handling:Protect from light and moisture.
 
Protocol:Simultaneous Fixation, Permeabilization, and Fluorescent Phallotoxin Staining
The phallotoxins appear to be stable for short periods in 4% formaldehyde fixation buffers.
This permits a rapid one-step fixation, permeabilization, and labeling procedure as follows.
 
Preparation Instructions
Solutions should be prepared fresh and protected from light when ever possible.
Solubility in water (0 °C): 0.5%; much more soluble in hot water; freely soluble in methanol, ethanol, butanol, and pyridine.
Solubility of product in methanol at the following concentration: Phalloidin-FITC: 0.5 mg/ml
 
Procedure
Stock solutions of phalloidin conjugates have been made in methanol or DMSO at 0.1–5 mg/ml. Final staining solutions in aqueous physiological buffers are in the concentration range of 0.1–100 µM with
corresponding incubation times of 15 minutes to 72hours.
The following procedure may be used as a guideline for staining cells:Waggoner, A. et al., Methods in Cell Biology, 30, 449 (1989)
 
  1. Cells are washed with phosphate buffered saline (PBS).
  2. Cells are fixed for 5 minutes in 3.7% formaldehyde solution in PBS. Then washed extensively in PBS.
  3. Cells may be dehydrated with acetone, permeabilized with 0.1% TRITON ® X-100 in PBS, and washed again in PBS.
  4. Cells are stained with a 50 μg/ml fluorescent phalloidin conjugate solution in PBS (containing 1% DMSO from the original stock   solution) for 40 minutes at room temperature.
  5. Wash several times with PBS to remove unbound phalloidin conjugate.
  6. Mount coverslips and view.
Excitation: 495 nm
Emission: 513 nm
 
Living Cells
Phallotoxins are usually not cell-permeant and have therefore not been used extensively with living cells. However, living cells have been labeled.
Pinocytosis appears to be the method of entry for some cells, although hepatocytes “avidly” take up the dye by an unknown mechanism.
In general, a larger amount of stain will be needed for staining living cells.
 
350-268 1
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350-268 1

Product Literature References

The PR-1 domain accounts for the anti-angiogenic activity of a cysteine-rich secretory protein member from the buccal glands of Lampetra japonica: D. Duan, et al.; Int. J. Biol. Macromol. (2017), Abstract;
Antimicrobial activity of tantalum oxide coatings decorated with Ag nanoparticles: H. Cao, et al.; J. Vac. Sci. Technol. A 34, 04C102 (2016), Application(s): Cell staining, Full Text
CD99 inhibits CD98-mediated β1 integrin signaling through SHP2-mediated FAK dephosphorylation: K.J. Lee, et al.; Exp. Cell Res. 336, 211 (2015), Application(s): Cell Culture, Abstract;
Effects of mechanical stretching on the morphology and cytoskeleton of vaginal fibroblasts from women with pelvic organ prolapse: S. Wang, et al.; Int. J. Mol. Sci. 16, 9406 (2015), Application(s): Cell Culture, Microscopy, Abstract; Full Text
The effect of hyaluronan on the motility of skin dermal fibroblasts in nanofibrous scaffolds: Y. Qian, et al.; Int. J. Biol. Macromol. 79, 133 (2015), Application(s): Immunofluorescence, Abstract;
F‑actin cytoskeleton reorganization is associated with hepatic stellate cell activation: X. Cui, et al.; Mol. Med. Rep. 9, 1641 (2014), Abstract; Full Text
Fluorescent phallotoxins as probes for filamentous actin: H. Faulstich, et al.; J. Muscle Res. Cell Motil. 9, 370 (1988), Abstract;
Preparation of tetramethylrhodaminyl-phalloidin and uptake of the toxin into short-term cultured hepatocytes by endocytosis: H. Faulstich, et al.; Exp. Cell. Res. 144, 73 (1983), Abstract;
Fluorescent phallotoxin, a tool for the visualization of cellular actin: E. Wulf, et al.; PNAS 76, 4498 (1979), Abstract;

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ALX-350-265-M001 1 mg 182.00 USD
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