Chemical Structure
(Molecular Diagram)
Fluor 488-Phalloidin Staining Reagent
Fluor 488-Phalloidin
Synonyms
Fluor 488-Phalloidin; N/A; Fluor 488-Phalloidin staining reagent
Samples
To label cellular F-actin
Assay Procedure
1. Prepare 100×AbFluor™ 488-Phalloidin DMSO stock solution: by adding 500 ?L of DMSO into the powder form vials. Then should be aliquoted and stored at -20°C. Protected from light and avoid freeze/thaw cycles.
2. Prepare 1×AbFluor™ 488-Phalloidin conjugate working solution: by adding 1 ?L of 100× Phalloidin conjugate DMSO solution to 99 uL of 1× PBS.
Note: Because the optimal staining concentration used for different cells may be different, the dilution ratio can be appropriately adjusted according to the actual staining effect.
3. Grow cells directly on a coverslip in 96 well dish. Incubate in a CO2 incubator at 37°C for at least 24 h before treatment.
4. Wash cells with PBS twice before fix cells with ice-cold 4% formaldehyde fixation for 15-30 min on the ice.Note: Methanol can damage actin during the fixation process. So, it is best to avoid fixatives containing any methanol. The preferred fixative is formaldehyde free of methanol.
5. Wash cells with PBS three times and cells were permeabilized with 0.1% Triton X-100 in PBS for 10 min at room temperature.
6. Wash cells with PBS three times and Add 100 uL/well (96-well plate) of AbFluor™ 488-Phalloidin staining solution into cells, and stain cells for 30 min at room temperature.
7. Wash cells with PBS twice and viewed and recorded using fluorescence microscope. The AbFluor™ 488-Phalloidin has good light stability and the sample can be imaged in PBS, but for best effect, it can be observed using an anti-fluorescence quencher.
2. Prepare 1×AbFluor™ 488-Phalloidin conjugate working solution: by adding 1 ?L of 100× Phalloidin conjugate DMSO solution to 99 uL of 1× PBS.
Note: Because the optimal staining concentration used for different cells may be different, the dilution ratio can be appropriately adjusted according to the actual staining effect.
3. Grow cells directly on a coverslip in 96 well dish. Incubate in a CO2 incubator at 37°C for at least 24 h before treatment.
4. Wash cells with PBS twice before fix cells with ice-cold 4% formaldehyde fixation for 15-30 min on the ice.Note: Methanol can damage actin during the fixation process. So, it is best to avoid fixatives containing any methanol. The preferred fixative is formaldehyde free of methanol.
5. Wash cells with PBS three times and cells were permeabilized with 0.1% Triton X-100 in PBS for 10 min at room temperature.
6. Wash cells with PBS three times and Add 100 uL/well (96-well plate) of AbFluor™ 488-Phalloidin staining solution into cells, and stain cells for 30 min at room temperature.
7. Wash cells with PBS twice and viewed and recorded using fluorescence microscope. The AbFluor™ 488-Phalloidin has good light stability and the sample can be imaged in PBS, but for best effect, it can be observed using an anti-fluorescence quencher.
Precautions
1. Please immediately centrifugal the product to the bottom of the tube before use, and then conduct the subsequent experiments.
2. Phalloidin is harmful to human body, please wear a laboratory coat and disposable gloves to operate, pay attention to appropriate protection.
3. Fluorescent dyes all have quenching problems, please try to avoid light to slow down the quenching.
2. Phalloidin is harmful to human body, please wear a laboratory coat and disposable gloves to operate, pay attention to appropriate protection.
3. Fluorescent dyes all have quenching problems, please try to avoid light to slow down the quenching.
Preparation and Storage
Store at -20°C for 12 months, protect from light.
Chemical Structure
(Molecular Diagram)
Chemical Structure
(Molecular Diagram)
Related Product Information for Fluor 488-Phalloidin staining reagent
Principle of the Assay: Phalloidin has the molecular formula C35H48N8O11S, molecular weight 788.87 and CAS number 17466-45-4. Phalloidin belongs to a class of toxins called phallotoxins, which are isolated from the death cap mushroom (Amanita phalloides). It is a bicyclic peptide that binds to F-actin specifically. Therefore, the distribution of F-actin can be very conveniently studied by using a fluorescent dye-labeled phalloidin. Inside the phalloidin, there is an unusual thioether bridge between cysteine and tryptophan, which can form an inner ring structure. When the pH is raised, the thioether is cleaved and the phalloidin loses its affinity for actin. AbFluor™ 488-Phalloidin selectively bound to F-actin, it is much higher photostability than the fluorescein-phalloidin conjugates. Phalloidin derivatives are convenient probes for labeling, identifying and quantitating F-actin in formaldehyde-fixed and permeabilized tissue sections, which can stain F-actin in cell cultures or cell-free experiments at nanomolar levels.
Product Categories/Family for Fluor 488-Phalloidin staining reagent
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Product Notes
The Fluor 488-Phalloidin (Catalog #AAA332505) is a Staining Reagent and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "Fluor 488-Phalloidin, Staining Reagent" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
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