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Phalloidin-iFluor 405 Reagent (ab176752) is one of a series of phalloidin conjugates that bind to actin filaments, also known as F-actin. Phalloidin-iFluor 405 can be easily detected with a fluorescent microscope at Ex/Em = 400/421 nm.
Phalloidin conjugates are convenient probes for labeling, identifying and quantifying animal or plant actin filaments in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments. They can also be used in paraffin-embedded samples that have been de-paraffinized.
Review our other popular phalloidin dye conjugates, including Phalloidin-iFluor 488, Phalloidin-iFluor 647, Phalloidin-iFluor 594, Phalloidin-iFluor 555, and Rhodamine Phalloidin, or search our website to see the rest of the range.
Staining fixed cell or tissue samples with phalloidin conjugates is very simple; it requires a single 20-90 min incubation with the phalloidin, followed by 3 short wash steps. Phalloidin staining can be combined with antibody-based staining by adding the phalloidin conjugate during either the primary or secondary antibody incubation step.
When used in unfixed samples, phalloidin binding leads to a decrease in the disassociation rate of actin subunits from the ends of actin filaments, essentially stabilizing actin filaments through the prevention of filament depolymerisation.
|Phalloidin-iFluor 405 Conjugate||1 x 300 tests|
Our Abpromise guarantee covers the use of ab176752 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|FM||Use at an assay dependent concentration.|
CytoPainter Phalloidin-iFluor 405 Reagent was used to stain F-actin in Bovine Foetal Aeorta Endothelial (BFA) cells. This was tested on triton x-100 permeabilised and non-permeabilised cells, phalloidin was prepared at 1 in 1000, diluted in 1% BSA in PBS. Cells were stained for 20 minutes. Coverslips with cells were then prepared and used for confocal microscopy. Nucleus was also stained with a nuclear stain that is excited by 633 laser. ctin staining worked in both permeabilised and unpermeabilised cells, although fluorescence was dimmer when compared to phalloidin-488 or phalloidin-568 stain, however this could be overcome perhaps by leaving the product on the cells for longer. Fluoresence also bleached relatively quickly, so it was important not to excite the fluorophore too strongly or for too long. Despite this, cells stained well, showing the same actin structures seen with other phalloidin stains and had the advantage in that the product could be used more dilute (1 in 1000) compared to other stains that are used at 1 in 25 dilution. I would recommend this product if 488 or 568 fluorophores could not be used.
Excitation and emission spectra of phalloidin-iFluor 405 reagent
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"