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Phalloidin Conjugates

A highly selective actin filament stain for fixed and permeabilized cells. Labeled phalloidins have similar affinity for both large and small filaments.

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Product Description

A toxin isolated from the deadly Amanita phalloides mushroom. It is a bicyclic peptide that binds specifically to F-actin. When conjugated to a fluorescent label, it will stain actin filaments in fixed and permeabilized cells.

  • Choice of 17 bright & photostable CF® Dyes, biotin, or other common fluorophores
  • Dye options for super-resolution and 2-photon imaging
  • Excellent signal to background with negligible non-specific staining

Note: Biotium also offers ActinBrite™ High Affinity Phalloidin Conjugates which were designed to preserve high phalloidin affinity for F-actin, resulting in bright staining that can be imaged more than one month after staining with minimal loss of signal or specificity.

Fluorescently-labeled phalloidins bind to F-actin with nanomolar affinity. Labeled phalloidins have similar affinity for both large and small filaments, binding in a stoichiometric ratio of about one phalloidin molecule per actin subunit in muscle and nonmuscle cells from various species of plants, animals and fungi. Different from antibodies, the binding affinity of phalloidin does not change significantly with actin among different species.

Fluorescently-labeled phalloidin is typically used to stain fixed and permeabilized cells. Labeled phalloidins are not cell-permeant and have therefore not been used extensively with living cells, however they can be loaded into live cells via cationic liposomes.

Phalloidin stock solutions are very stable. But binding of fluorescent phalloidins to cells is more labile than other probes, such as antibodies, and staining can become non-specific or lost over time. For best results, store phalloidin-stained samples in a suitable mounting medium at 4°C, protected from light. For certain phalloidin conjugates, especially CF®405M, CF®647, and CF®680, we recommend imaging immediately or shortly after staining. Staining with our other CF® Dye phalloidins usually is stable for up to a week when specimens are stored at 4°C, protected from light.

For stable F-actin staining, Biotium recommends ActinBrite™ High Affinity Phalloidin Conjugates which were designed to preserve strong F-actin binding. With ActinBrite™, samples can be imaged after for a month or more (depending on the conjugate and mounting method)—making delayed imaging easier and more dependable.

Superior CF® Dyes

Biotium’s next-generation CF® Dyes were designed to be highly water-soluble with advantages in brightness and photostability compared to Alexa Fluor®, DyLight®, and other fluorescent dyes. Learn more about CF® Dyes.

Note: Conjugates of blue-fluorescent dyes like CF®350, CF®405S and CF®405M are not recommended for detecting low abundance targets and may be challenging to use in tissue specimens. Blue dyes have lower fluorescence and photostability, and cells and tissue have high autofluorescence in blue wavelengths, resulting in lower signal to noise compared to other colors.

Super-Resolution Microscopy

CF®647 and CF®680 phalloidins are recommended mainly for STORM applications. Certain CF® Dyes are compatible with various super resolution imaging techniques. The superior brightness, photostability, and photochemical switching properties of certain CF® Dyes are ideal for 3-D SIM, 3-D STORM, and other super-resolution and single-molecule imaging approaches. Learn more about CF® Dyes for super-resolution microscopy.

Phalloidin Conjugates

ProductConjugationEx/EmSizeCatalog No.Purchase
Biotin-XX PhalloidinBiotin-XXN/A100 U00028Purchase 00028
CF®350 PhalloidinCF®350347/448 nm50 U00049-TPurchase 00049-T
300 U00049Purchase 00049
CF®405M PhalloidinCF®405M408/452 nm50 U00034-TPurchase 00034-T
300 U00034Purchase 00034
CF®430 PhalloidinCF®430426/498 nm50 U00054-TPurchase 00054-T
300 U00054Purchase 00054
CF®440 PhalloidinCF®440440/515 nm50 U00055-TPurchase 00055-T
300 U00055Purchase 00055
CF®488A PhalloidinCF®488A490/515 nm50 U00042-TPurchase 00042-T
300 U00042Purchase 00042
CF®532 PhalloidinCF®532527/558 nm50 U00051-TPurchase 00051-T
300 U00051Purchase 00051
CF®543 PhalloidinCF®543541/560 nm50 U00043-TPurchase 00043-T
300 U00043Purchase 00043
CF®568 PhalloidinCF®568562/583 nm50 U00044-TPurchase 00044-T
300 U00044Purchase 00044
CF®583R PhalloidinCF®583R586/609 nm50 U00064-TPurchase 00064-T
300 U00064Purchase 00064
CF®594 PhalloidinCF®594593/614 nm50 U00045-TPurchase 00045-T
300 U00045Purchase 00045
CF®633 PhalloidinCF®633630/650 nm50 U00046-TPurchase 00046-T
300 U00046Purchase 00046
CF®640R PhalloidinCF®640R642/662 nm50 U00050-TPurchase 00050-T
300 U00050Purchase 00050
CF®647 Phalloidin*CF®647650/665 nm50 U00041-TPurchase 00041-T*
300 U00041Purchase 00041*
CF®660C PhalloidinCF®660C667/685 nm50 U00052-TPurchase 00052-T
300 U00052Purchase 00052
CF®660R PhalloidinCF®660R663/682 nm50 U00047-TPurchase 00047-T
300 U00047Purchase 00047
CF®680 Phalloidin*CF®680681/698 nm50 U00053-TPurchase 00053-T*
300 U00053Purchase 00053*
CF®680R PhalloidinCF®680R680/701 nm50 U00048-TPurchase 00048-T
300 U00048Purchase 00048
Fluorescein PhalloidinFluorescein496/516 nm300 U00030Purchase 00030
Rhodamine PhalloidinRhodamine540/565 nm300 U00027Purchase 00027
Rhodamine 110 PhalloidinRhodamine 110502/524 nm300 U00032Purchase 00032
Sulforhodamine 101 (Texas Red®) PhalloidinTexas Red®591/608 nm300 U00033Purchase 00033
*CF®647 and CF®680 phalloidins are recommended for STORM applications, but due to the instability of staining with these conjugates, we do not recommend using them for other microscopy applications. For other applications requiring far-red phalloidins, we recommend CF®633, CF®640R, or CF®680R phalloidins, which have more stable binding.

Note: CF®555 Phalloidin (00040, 00040-T) has been discontinued. We recommend CF®568 Phalloidin as a replacement for conventional microscopy, or CF®583R Phalloidin as a replacement for STORM.

CF is a registered trademark of Biotium, Inc. Alexa Fluor, Texas Red, and DyLight are registered trademarks of Thermo Fisher Scientific.

Product Attributes

Size
100 U, 50 U, 300 U
Conjugation
CF®350, CF®405M, CF®430, CF®440, CF®488A, CF®532, CF®543, CF®555, CF®568, CF®583R, CF®594, CF®633, CF®640R, CF®647, CF®660C, CF®660R, CF®680, CF®680R, Fluorescein (FITC), Sulforhodamine 101 (Texas Red®), Biotin-XX, Rhodamine, Rhodamine 110
Probe cellular localization
Cytoskeleton, F-Actin
For live or fixed cells
For fixed cells
Assay type/options
Tissue staining
Detection method/readout
Fluorescence microscopy
Cell permeability
Membrane impermeant
Fixation options
Fix before staining (formaldehyde), Permeabilize before staining
Toxin
Phalloidin
Colors
Blue, Green, Orange, Red, Far-red, Near-infrared

Documents, Protocols, SDS and COA

References

Download a list of CF® dye Phalloidin references.

FAQs

Product shipping, storage, shelf life, & solubility

Bioscience kits
The guaranteed shelf life from date of receipt for bioscience kits is listed on the product information sheet. Some kits have an expiration date printed on the kit box label, this is the guaranteed shelf life date calculated from the day that the product shipped from our facility. Kits often are functional for significantly longer than the guaranteed shelf life. If you have an older kit in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the kit still works for your application before processing a large number of samples or precious samples.

Antibodies and other conjugates
The guaranteed shelf life from date of receipt for antibodies and conjugates is listed on the product information sheet. Antibodies and other conjugates often are functional for significantly longer than the guaranteed shelf life. If you have an older conjugate in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples.

For lyophilized antibodies, we recommend reconstituting the antibody with glycerol and antimicrobial preservative like sodium azide for the longest shelf life (note that sodium azide is not compatible with HRP-conjugates).

Chemicals, dyes, and gel stains
Biotium guarantees the stability of chemicals, dyes, and gel stains for at least a year from the date you receive the product. However, the majority of these products are highly stable for many years, as long as they are stored as recommended. Storage conditions can be found on the product information sheet or product safety and data sheet, material safety data sheet, and on the product label. Fluorescent compounds should be protected from light for long term storage.

If you have a Biotium compound that has been in storage for longer than one year that you wish to use, we recommend performing a small scale positive control experiment to confirm that the compound still works for your application before processing a large number of samples or precious samples.

Expiration date based on date of manufacture (DOM)
If your institution requires you to document expiration date based on date of manufacture for reagents, please contact techsupport@biotium.com for assistance.

Chemical products with special stability considerations:

Esters

Ester compounds include the following:

  • Succinimidyl esters (SE, also known as NHS esters), such as our amine-reactive dyes
  • Acetoxymethyl esters (AM esters), such as our membrane-permeable ion indicator dyes
  • Diacetate-modified dyes, like ViaFluor™ 405, CFDA, and CFDA-SE cell viability/cell proliferation dyes

Ester dyes are stable in solid form as long as they are protected from light and moisture. Esters are not stable in aqueous solution. Concentrated stock solutions should be prepared in anhydrous DMSO (see Biotium catalog no. 90082). Stock solutions in anhydrous DMSO can be stored desiccated at -20°C for one month or longer. Esters should be diluted in aqueous solution immediately before use. Succinimidyl esters (SE) should be dissolved in a solution that is free of amine-containing compounds like Tris, glycine, or protein, which will react with the SE functional group. AM esters and diacetate compounds should be dissolved in a solution that is free of serum, because serum could contain esterases that would hydrolyze the compound.

A note on CF® Dye succinimidyl ester stability

Succinimidyl esters (SE) are generally susceptible to hydrolysis, which can result in lower labeling efficiency. Many commercially available fluorescent dyes used for life science research are heavily sulfonated dyes which makes them particularly hygroscopic, worsening the hydrolysis problem. In addition, for several commercially available SE reactive dyes, the SE group is derived from an aromatic carboxylic acid, while the SE group in all of Biotium’s CF® Dyes is prepared from an aliphatic carboxylic acid. This structural difference reduces the susceptibility of CF® Dye SE reactive groups to hydrolysis, resulting in relatively stable reactive dyes with consistently higher labeling efficiency compared to other SE derivatives of other fluorescent dyes.

Maleimides, MTS and thiosulfate dyes
Like the succinimidyl ester dyes, these dyes are also susceptible to hydrolysis, although generally to a much lower degree. Thus, for long term storage, anhydrous DMSO is recommended for making stock solutions.

Other reactive dyes
Amines, aminooxy (also known as oxylamine), hydrazide, azide, alkyne, BCN, and tyramide reactive dyes, as well as dye free acids, are generally stable in aqueous solution when stored at -20°C for 6-12 months or longer, as long as no compounds are present that may react with the dye’s functional group. See the product information sheets for specific reactive dyes more information.

Coelenterazines and D-luciferin

Coelenterazines are stable in solid form when stored as recommended; they are not stable in aqueous solution. Concentrated coelenterazine stock solutions (typically 1-100 mg/mL) should be prepared in ethanol or methanol; do not use DMSO or DMF to dissolve coelenterazines, because these solvents will oxidize the compounds. Ethanol or methanol stocks of coelenterazine can be stored at -20°C or below for six months or longer; alcohol stocks may evaporate during storage, so use tightly sealing screw cap vials and wrap the vials with Parafilm for long term storage. Propylene glycol also can be used as a solvent to minimize evaporation. If the solvent evaporates, the coelenterazine will still be present in the vial, so note the volume in the vial prior to storage so that you can adjust the solvent volume to correct for evaporation if needed. Prepare working solutions in aqueous buffers immediately before use. Coelenterazines are stable for up to five hours in aqueous solution.

Aquaphile™ coelenterazines are water soluble formulations of coelenterazines. They are stable in solid form when stored as recommended. Aquaphile™ coelenterazines should be dissolved in aqueous solution immediately before use. They are stable for up to five hours in aqueous solution.

Note that coelenterazines are predominantly yellow solids, but may contain dark red or brown flecks. This does not affect product stability or performance. If your coelenterazine is uniformly brown, then it is oxidized and needs to be replaced.

D-luciferin is stable in solid form and as a concentrated stock solution when stored as recommended; it is not stable at dilute working concentrations in aqueous solution. Prepare concentrated D-luciferin stock solutions (typically 1-100 mg/mL) in water, and store in aliquots at -20°C or below for six months or longer. Prepare working solutions immediately before use.

For dyes or reagents that are supplied lyophilized (as solids), it is hard to compare quantities based on appearance of the dye in the tube, because during the lyophilization process the dye can dry down in different ways, either spread out all over the tube, clumped together, or coating the sides or bottom of the tube. Centrifugation of the tube may not help in collecting the dye solid to the bottom of the tube as this generally works for solutions. However, lyophilized solids are packaged based on highly accurate absorbance measurement of the reagent solution prior to drying, so the vial will contain the correct amount of dye.

Biotium ships all antibodies (primary, secondary and conjugates) at room temperature. We guarantee their quality and performance under these conditions based upon our stability testing. Antibodies were subjected to accelerated stability testing by storing them at various temperatures (4°C, room temperature, or 37°C) for 1 week to mimic simulated shipping conditions and tested in immunostaining experiments. All antibodies showed the expected brightness and specificity, even after storage at sub-optimal temperatures for a week or longer. You can also download our Product Storage Statement here.

In line with our goal to be more environmentally friendly by reducing the use of excess packaging, and lowering shipping costs for our customers, products that have passed our stability testing are shipped at room temperature.

Once you have received the antibody vial, please follow the long-term storage instructions on the product information (PI) sheet.

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