Phalloidin
Phalloidin kdorfman Thu, 08/13/2015 - 21:03General protocol:
Stock Solution: Add 1.5 mL MeOH to vial (~6.6µM)
Aliquots
- 5 µL stock per 0.5 mL tube
- Label: Add 200 µL PBS (or PBS 1% BSA), use 50µL per coverslip
Fix cells with formaldehyde
Procedure:
- 50 uL phalloidin on parafilm in humid chamber
- coverslip cell side down onto drop
- 15 - 20 minutes at room temp
- rinse repeatedly in PBS-Tween-azide
- blot the corner on a kimwipe
- Mount on a drop of mounting medium on a slide
FITC phalloidin
FITC phalloidin kdorfman Fri, 06/01/2018 - 18:40Thermo Fisher F432
From the manufacturer:
Fluorescein phalloidin is a high-affinity F-actin probe conjugated to the green fluorescent dye, fluorescein (FITC).
- Selectively stains F-actin
- Excitation/Emission: 496/516 nm
- Superior to antibody staining
- Optimal for fixed and permeabilized samples
10 and 20 uL aliquots
In histology freezer
Red phalloidins
Red phalloidins kdorfman Fri, 06/01/2018 - 18:42Alexa Fluor 568 Phalloidin
Alexa Fluor 568 Phalloidin kdorfman Fri, 06/01/2018 - 18:41Alexa Fluor 568 Phalloidin
Fisher or Invitrogen A 12380
Binds to F-Actin
Ex/Em 578/600
Stock Solution: Add 1.5 mL MeOH to vial (~6.6µM)
Aliquots
- 8 µL stock per 0.5 mL tube
- Label: Add 400 µL PBS (or PBS 1% BSA), use 50µL per coverslip
Fix cells with formaldehyde
Procedure:
- 50 uL phalloidin on parafilm in humid chamber
- coverslip cell side down onto drop
- 15 - 20 minutes at room temp
- rinse repeatedly in PBS-Tween-azide
- blot the corner on a kimwipe
- Mount on a drop of mounting medium on a slide
In histology and bioimaging freezers
Rhodamine Phalloidin
Rhodamine Phalloidin kdorfman Fri, 06/01/2018 - 18:35Thermo Fisher R415
From the manufacturer:
Rhodamine phalloidin is a high-affinity F-actin probe conjugated to the red-orange fluorescent dye, tetramethylrhodamine (TRITC).
- Selectively stains F-actin
- Excitiation/Emission: 540/565 nm
- Superior to antibody staining
- Optimal for fixed and permeabilized samples
- Very widely cited fluorescent phalloidin conjugate
In 262A freezer
Texas Red-X Phalloidin
Texas Red-X Phalloidin kdorfman Fri, 06/01/2018 - 18:30Thermo Fisher T7471
From the manufacturer:
Texas Red®-X phalloidin is a high-affinity F-actin probe conjugated to our bright, photostable, red fluorescent Texas Red®-X dye.
- Selectively stains F-actin
- Outstanding fluorescence performance
- Excitation/Emission: 591/608 nm
- Superior to antibody staining
- Optimal for fixed and permeabilized samples
In histology freezer