Invitrogen™ eBioscience™ Phalloidin eFluor™ 660

Catalog No.	Quantity
50-112-4427	300 tests

Catalog No. 50-112-4427 Supplier Invitrogen™ Supplier No. 50655905

Description

Can be used to label cells of multiple species via immunohistochemical staining. eBioscience™ Phalloidin eFluor™ 660 Solution is supplied as a lyophilized solid which, when reconstituted, forms a 1000x stock solution that can be used to prepare a 1x working solution.

Phalloidin is a peptide toxin isolated from the Amanita phalloides mushroom. Phalloidin selectively binds F-actin and functions to stablilize actin polymers by inhibiting the dissociation of actin monomers from the filament ends. Because F-actin polymers compose the cytoskeleton of almost all eukaryotic cells, phalloidin conjugates can be useful probes for visualizing the structure of live or fixed cells using microscopy. Phalloidin eFluor 660 can be used to label cells of multiple species.

Phalloidin eFluor 660 is supplied as a lyophilized solid and should be prepared by reconstitution with 30μL high-quality anhydrous DMSO to give a 1000X stock solution. The phalloidin DMSO stock solution should be aliquoted, protected from light and moisture, and stored at -20°C. To prepare a 1X working solution of phalloidin conjugate, add 1μL of 1000X phalloidin conjugate DMSO solution to 1mL of PBS containing 1% BSA.

Phalloidin is a peptide toxin isolated from the Amanita phalloides mushroom. Phalloidin selectively binds F-actin and functions to stabilize actin polymers by inhibiting the dissociation of actin monomers from the filament ends. Because F-actin polymers compose the cytoskeleton of almost all eukaryotic cells, phalloidin conjugates can be useful probes for visualizing the structure of live or fixed cells using microscopy.

• Phalloidin eFluor™ 660 is supplied as a lyophilized solid
• To prepare the 1000X stock solution, the material should be mixed with 30μL high-quality anhydrous DMSO
• The resulting phalloidin DMSO stock should be aliquoted, protected from light and moisture, and stored at −20°C
• To prepare a 1X working solution of phalloidin conjugate, add 1μL of 1000X phalloidin DMSO stock solution to 1mL of PBS containing 1% BSA

Specifications

• Excitation Wavelength Range: 633-647/668
• Dye Type: eFluor 660
• No. of Tests: 300 tests
• Quantity: 300 tests
• Product Line: eBioscience
• Label Type: eFluor
• Product Type: Phalloidin
• SubCellular Localization: Cytoskeleton

Frequently Asked Questions (FAQs)

Can I combine Click-iT or Click-iT Plus reactions with phalloidin conjugates used for actin staining?

We do not recommend using phalloidin conjugates for staining actin in combination with traditional Click-iT or Click-iT Plus reactions since phalloidin is extremely sensitive to the presence of copper.

For staining actin in combination with traditional Click-iT or Click-iT Plus reactions, we recommend using anti-α-actin antibodies for staining actin in the cytoskeleton. You can find a list of our actin antibodies here.

Another option would be to use the Click-iT Plus Alexa Fluor Picolyl Azide Toolkit (Cat. Nos. C10641, C10642, C10643). These Click-iT Plus toolkits provide Copper and Copper protectant separately which makes it easier to titrate the copper concentration to obtain optimal labeling with minimal copper-mediated damage. You may need to optimize the click reaction with the lowest possible concentration of copper and then perform the phalloidin staining.

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov

For Research Use Only. Not for use in diagnostic procedures.