Phalloidin Conjugates For Staining Actin
We provide a number of Invitrogen Molecular Probes fluorescent and biotinylated derivatives of phalloidin that deliver significant advantages over antibodies for actin labeling. The binding properties of phalloidin conjugates do not change appreciably with actin from different species, including plants and animals. In addition, they exhibit negligible nonspecific staining, so the contrast between stained and unstained areas is high.
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Phalloidin is a bicyclic peptide that belongs to a family of toxins isolated from the deadly Amanita phalloides “death cap” mushroom and is commonly used in imaging applications to selectively label F-actin in fixed cells, permeabilized cells, and cell-free experiments. Labeled phalloidin conjugates have similar affinity for both large and small filaments and bind in a stoichiometric ratio of about one phallotoxin per actin subunit in both muscle and nonmuscle cells; they reportedly do not bind to monomeric G-actin, unlike some antibodies against actin.
Alexa Fluor and Alexa Fluor Plus phalloidin conjugates
Fluorescent Alexa Fluor and Alexa Fluor Plus dye conjugates of phalloidin are widely used F-actin stains, with color choices across the full spectral range. These phalloidin conjugates provide researchers with fluorescent probes that are designed to be superior in brightness and photostability to all other spectrally similar conjugates tested.
Fixed, permeabilized bovine pulmonary artery endothelial cells visualized using components of the Invitrogen SelectFX Nuclear Labeling Kit and Alexa Fluor phalloidin conjugates. Nuclei and F-actin were stained, respectively, with (top left) DAPI and Invitrogen Alexa Fluor 680 phalloidin, (top right) Invitrogen SYTOX Green dye and Invitrogen Alexa Fluor 568 phalloidin, (bottom left) 7-AAD and Invitrogen Alexa Fluor 488 phalloidin, and (bottom right) Invitrogen TO-PRO 3 dye and Invitrogen Alexa Fluor 350 phalloidin.
Spectral characteristics of Invitrogen phalloidin probes
|Actin-selective probe||Ex/Em*||~ MW||Cat. No.|
|Alexa Fluor 350 Phalloidin||346/442||1100||A22281|
|Alexa Fluor Plus 405 Phalloidin||405/450||1010||A30104|
|Alexa Fluor 488 Phalloidin||496/519||1180||A12379|
|Oregon Green 488 phalloidin||496/520||1180||O7466|
|Alexa Fluor 555 Phalloidin||555/565||1910||A34055|
|BODIPY 558/568 phalloidin||558/569||1115||B3475|
|Alexa Fluor 594 Phalloidin||590/617||1620||A12381|
|Texas Red-X phalloidin||591/608||1490||T7471|
|Alexa Fluor 647 Phalloidin||650/665||1950||A22287|
|Alexa Fluor 680 Phalloidin||679/702||1850||A22286|
|Alexa Fluor Plus 750 Phalloidin||758/784||2122||A30105|
|* Excitation (Ex) and emission (Em) maxima, in nm.|