Mast Cell Degranulating (MCD) Peptide

Phalloidin is a mushroom-derived toxin which can be used to label F-actin of the cytoskeleton with fluorochrome(λex=495 nm, λem=520 nm). Sequence: Ala-{d-Thr}-Cys-{Hyp}-Ala-Trp-Leu (Disulfide bridge: Cys3-Cys6);A-{d-Thr}-C-{Hyp}-AWL (Disulfide bridge: Cys3-Cys6).

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Peptides

Phalloidin staining will not work with methanol-fixed cells, probably because of the disruption of actin filament integrity, but it will work with cells fixed in 0.2% glutaraldehyde in PBS. For fluorescein isothiocyanate (FITC)-Phalloidin (or rhodamine isothiocyanate [RITC]-Phalloidin), the excitation wavelength maximum will be near 495 nm (or 550 nm) and the emission wavelength maximum will be ~520 nm based on H2O at pH 8.0 as the solvent (575 nm with methanol as the solvent). The emission spectrum of fluorescein can depend on the environmental pH and/or ionic strength[1].

Prepare a 1:200 dilution of the stock solution (300 units/mL) of fluorescein Phalloidin or rhodamine Phalloidin using PBS. Aspirate the cell medium from the cells grown on glass coverslips and rinse the cells three times with PBS. Fix the cells for 10 min in the 3.7% formaldehyde solution. Rinse the fixed cells three times for 5 min each in PBS. Permeabilize the cells for 5 min in the 0.2% Triton X-100 solution. Rinse the fixed cells three times with PBS. Label the cells with the fluorescein or rhodamine Phalloidin for 5 to 10 min at room temperature. Rinse the cells three times in PBS for 5 min each time. Mount the cell and view the cells using either the fluorescein or rhodamine filter set, depending on the probe[1].

[1]. Chazotte B. Labeling cytoskeletal F-actin with rhodamine phalloidin or fluorescein phalloidin for imaging. Cold Spring Harb Protoc. 2010 May;2010(5):pdb.prot4947.

Captisol | Cyclosporin A | H2DCFDA | 0MPTP hydrochloride | GW4869 | Etomoxir | TD139 | Mitoquinone mesylate | GSK2795039 | JC-1 | BAPTA-AM | AP 20187 | Setanaxib (GKT137831) | D-Luciferin | Crotaline

MOLECULAR FORMULA :

C35-H48-N8-O11-S

MOLECULAR WEIGHT :

788.97

HEALTH HAZARD DATA

ACUTE TOXICITY DATA

TYPE OF TEST :

LDLo - Lowest published lethal dose

ROUTE OF EXPOSURE :

Intraperitoneal

SPECIES OBSERVED :

Rodent - rat

DOSE/DURATION :

1 mg/kg

TOXIC EFFECTS :

Details of toxic effects not reported other than lethal dose value

TYPE OF TEST :

LD50 - Lethal dose, 50 percent kill

ROUTE OF EXPOSURE :

Intraperitoneal

SPECIES OBSERVED :

Rodent - mouse

DOSE/DURATION :

2 mg/kg

TOXIC EFFECTS :

Details of toxic effects not reported other than lethal dose value

TYPE OF TEST :

LDLo - Lowest published lethal dose

ROUTE OF EXPOSURE :

Intravenous

SPECIES OBSERVED :

Rodent - mouse

DOSE/DURATION :

6600 ug/kg

TOXIC EFFECTS :

Behavioral - muscle weakness Liver - fatty liver degeneration

TYPE OF TEST :

LD50 - Lethal dose, 50 percent kill

ROUTE OF EXPOSURE :

Unreported

SPECIES OBSERVED :

Rodent - mouse

DOSE/DURATION :

2 mg/kg

TOXIC EFFECTS :

Details of toxic effects not reported other than lethal dose value

TYPE OF TEST :

LD - Lethal dose

ROUTE OF EXPOSURE :

Intravenous

SPECIES OBSERVED :

Mammal - dog

DOSE/DURATION :

>10 mg/kg

TOXIC EFFECTS :

Details of toxic effects not reported other than lethal dose value

TYPE OF TEST :

LDLo - Lowest published lethal dose

ROUTE OF EXPOSURE :

Oral

SPECIES OBSERVED :

Mammal - species unspecified

DOSE/DURATION :

1000 ug/kg

TOXIC EFFECTS :

Details of toxic effects not reported other than lethal dose value

MUTATION DATA

TYPE OF TEST :

DNA inhibition

TEST SYSTEM :

Rodent - rat Liver

DOSE/DURATION :

100 nmol/L

REFERENCE :

TOXIA6 Toxicon. (Pergamon Press Ltd., Headington Hill Hall, Oxford OX3 OBW, UK) V.1- 1962- Volume(issue)/page/year: 25,1265,1987