What is Phalloidin stain?
What is Phalloidin stain?
Phalloidin is a highly selective peptide that is used for staining filamentous actin (F-actin) in formaldehyde-fixed and permeabilized tissue sections, cell-cultures and cell-free experiments. Phalloidin is commonly labeled with organic fluorophores such as iFluor™ dyes to facilitate visualization.
Is Phalloidin a fluorophore?
Phalloidin, a toxin isolated from the death cap mushroom, Amanita phalloides, binds to filamentous actin with high affinity, and this has made fluorophore-conjugated phalloidin a useful tool in cellular imaging.
What is Phalloidin FITC?
Phalloidin-FITC is a green fluorescent cytoskeleton stain. Binds and labels F-actin. For most fluorescent imaging applications, cells should be fixed and permeablilized. Excitation/emission maxima λ ~496/516 nm.
Why do we use phalloidin?
Phalloidin is a highly selective bicyclic peptide used for staining actin filaments (also known as F-actin). It binds to all variants of actin filaments in many different species of animals and plants.
How do you stain with phalloidin?
Phalloidin Staining Protocol
- Reconstitute phalloidin according to manufacturer’s directions.
- Fix cells in the collagen gels with 3.7% (v/v) paraformaldehyde 10 min at room temperature.
- Rinse 3 times in PBS.
- Permeabilize cells with 0.1 -0.5% (v/v) Triton X-100 for 10 min at room temperature.
- Rinse 3 times in PBS.
What does phalloidin do to liver cells?
Once inside the liver, phalloidin binds F-actin, preventing its depolymerization. It takes time for this process to destroy the liver cells. The kidneys can also take up phalloidin, but not as effectively as the liver. Here, phalloidin causes nephrosis.
Does phalloidin stabilize actin?
The cyclic peptide phalloidin binds and stabilizes actin filaments. It is widely used in studies of actin filament assembly, including analysis of branch formation by Arp2/3 complex, but its influence on the branching reaction has not been considered.
Where is actin produced?
The protein actin is abundant in all eukaryotic cells. It was first discovered in skeletal muscle, where actin filaments slide along filaments of another protein called myosin to make the cells contract.
What is F-actin staining?
Description F-Actin Stain is an easy-to-use probe-based solution for visualizing filamentous actin structures in fixed mammalian cells by fluorescence microscopy. F-actin is a major component of the cytoskeleton and is involved in fundamental cellular processes, such as cell division, morphogenesis, and migration.
What is SiR actin?
SiR-actin is fluorogenic, cell permeable and highly specific for F-actin. Sir-actin stains endogenous F-actin without the need for genetic manipulation or overexpression. Its emission in the far red minimizes phototoxicity and sample autofluorescence.
Why is phalloidin toxic?
The only apparent side effect of phalloidin poisoning is extreme hunger. This is because phalloidin is only taken up by the liver via bile salt membrane transport proteins. Once inside the liver, phalloidin binds F-actin, preventing its depolymerization. It takes time for this process to destroy the liver cells.
How do you dilute phalloidin?
Wash cells once with PBS. 8. For fluorescent phalloidin, dilute 5 μL methanolic stock solution of the phalloidin of your choice into 200 μL PBS with 1% BSA for each cover slip or chamber to be stained.
