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Invitrogen™ eBioscience™ Phalloidin eFluor™ 570
Description
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™ 570 can be used to label cells of multiple species.
Phalloidin eFluor™ 570 is supplied as a frozen liquid. To prepare the 1000X stock solution, thaw the vial, add 30 μL high-quality anhydrous DMSO, mix and briefly spin down. The resulting solution (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 1 μL of 1000X phalloidin DMSO stock solution to 1 mL of PBS containing 1% BSA.
Reported Applications
- Immunocytochemistry
- Immunohistochemical Staining
- Microscopy
Specifications
Specifications
| Excitation Wavelength Range | 555/570 nm |
| Dye Type | eFluor 570 |
| No. of Tests | 300 Tests |
| Quantity | 300 tests |
| Product Line | eBioscience |
| Label Type | eFluor |
| Product Type | Phalloidin |
| Sub Cellular Localization | Cytoskeleton |
Frequently Asked Questions (FAQs)
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.
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.
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).
The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.
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.
For Research Use Only. Not for use in diagnostic procedures.
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