| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Fluorescent dye
|
|---|---|
| ln Vitro |
1.1 Stock solution preparation
Prepare a 1 mM stock solution using DMSO as the solvent. 1.2 Working solution preparation Dilute the stock solution with pre-warmed serum-free medium or PBS buffer to a final concentration of 200-1000 nM. Note: Adjust the working solution concentration according to experimental requirements, and prepare fresh before use. 2.1 Cell pretreatment For suspension cells: Collect cells by centrifugation, then wash twice with PBS (5 minutes each time). For adherent cells: Remove culture medium, digest cells with trypsin, centrifuge to remove supernatant, then wash twice with PBS (5 minutes each time). 2.2 Staining procedure Add 1 mL of prepared Phalloidin-TRITC working solution and incubate at room temperature in the dark for 5-10 minutes. 2.3 Post-treatment steps Centrifuge at 400g for 3-4 minutes at 4°C, then discard the staining solution. Wash cells twice with PBS (5 minutes each time). Finally, resuspend cells in 1 mL of serum-free medium or PBS. 2.4 Observation and detection Observe and analyze the stained cells using a fluorescence microscope. |
| Cell Assay |
Phalloidin, the toxic drug from the mushroom Amanita phalloides, was injected into the cytoplasm of tissue culture cells and the changes in intracellular actin distribution were followed by immunofluorescence microscopy with actin antibody. At low concentrations, phalloidin recruits the non- or less highly polymerized forms of cytoplasmic actin into stable "islands" of aggregated actin polymers and does not interfere with the preexisting thick bundles of microfilaments (stress fibers). Differential focusing shows that these islands of phalloidin-induced actin polymers occur at a level in the cytoplasm that is above the submembranous bundles of microfilaments present on the adhesive side of the cells. The pattern of cytoplasmic microtubules remains unaffected by the injection of phalloidin; however, filamin, a protein usually associated with actin in the cytoplasm, is also recruited into the islands. At higher phalloidin concentrations, contraction of the cell is observed. These results are discussed in the light of previous biochemical studies by Wieland and Faulstich and their coworkers [for a review see Wieland, T. (1977) Naturwissenschaften 64, 303-309] on the in vitro interaction of phalloidin with muscle actin, which have documented that phalloidin reacts stoichiometrically with actin, promotes actin polymerization, and stabilizes actin polymers. In addition, we show that microinjection of phalloidin interferes in a concentration-dependent manner with cell locomotion and cell growth. These results indicate that a well-balanced controlled reversible equilibrium between different polymerization states of actin may be a necessary requirement for cell locomotion and may also influence other cellular functions such as growth [2].
|
| References | |
| Additional Infomation |
Tetramethylrhodamine phalloidin is a tetramethylrhodium dye coupled to the bicyclic peptide phalloidin via a thiourea bond. It is a fluorescent dye. Its function is related to that of phalloidin and tetramethylrhodamine.
|
| Molecular Formula |
C60H70N12O13S2
|
|---|---|
| Molecular Weight |
1231.400411129
|
| Exact Mass |
1230.462
|
| CAS # |
915013-10-4
|
| PubChem CID |
44140594
|
| Appearance |
Purple to purplish red solid powder
|
| LogP |
2.3
|
| Hydrogen Bond Donor Count |
12
|
| Hydrogen Bond Acceptor Count |
17
|
| Rotatable Bond Count |
8
|
| Heavy Atom Count |
87
|
| Complexity |
2780
|
| Defined Atom Stereocenter Count |
10
|
| SMILES |
N(C)(C1=CC2=[O+]C3C=C(N(C)C)C=CC=3C(C3C=CC(NC(=S)NC[C@@](O)(C)C[C@@H]4NC(=O)[C@]5(NC([C@H](C)NC(=O)C6C[C@@H](CN6C(C(CSC6NC7C(=CC=CC=7)C=6C5)NC(=O)[C@@]([H])([C@@H](O)C)NC(=O)[C@H](C)NC4=O)=O)O)=O)[H])=CC=3C([O-])=O)=C2C=C1)C
|
| InChi Key |
VXNOAAMNRGBZOQ-RUTQKCGOSA-N
|
| InChi Code |
InChI=1S/C60H70N12O13S2/c1-28-50(75)65-42-23-39-35-11-9-10-12-41(35)68-56(39)87-26-44(57(81)72-25-34(74)22-45(72)54(79)63-28)67-55(80)49(30(3)73)69-51(76)29(2)62-53(78)43(66-52(42)77)24-60(4,84)27-61-59(86)64-31-13-16-36(40(19-31)58(82)83)48-37-17-14-32(70(5)6)20-46(37)85-47-21-33(71(7)8)15-18-38(47)48/h9-21,28-30,34,42-45,49,68,73-74,84H,22-27H2,1-8H3,(H,61,86)(H,62,78)(H,63,79)(H,65,75)(H,66,77)(H,67,80)(H,69,76)(H,82,83)/b64-31+/t28-,29-,30-,34-,42-,43-,44-,45-,49+,60+/m0/s1
|
| Chemical Name |
(3E)-6-[3,6-bis(dimethylamino)xanthen-9-ylidene]-3-[[(2R)-2-hydroxy-3-[(1S,14R,18S,20S,23S,28S,31S,34R)-18-hydroxy-34-[(1S)-1-hydroxyethyl]-23,31-dimethyl-15,21,24,26,29,32,35-heptaoxo-12-thia-10,16,22,25,27,30,33,36-octazapentacyclo[12.11.11.03,11.04,9.016,20]hexatriaconta-3(11),4,6,8-tetraen-28-yl]-2-methylpropyl]carbamothioylimino]cyclohexa-1,4-diene-1-carboxylic acid
|
| Synonyms |
Rhodamine-phalloidin; 915013-10-4; starbld0003102; CHEBI:52313
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.8121 mL | 4.0604 mL | 8.1208 mL | |
| 5 mM | 0.1624 mL | 0.8121 mL | 1.6242 mL | |
| 10 mM | 0.0812 mL | 0.4060 mL | 0.8121 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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