| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
Rhod-2 AM targets intracellular calcium ions (Ca²⁺). The compound does not target a specific protein or enzyme but rather serves as a fluorescent calcium indicator. The AM ester form is cell-permeable and is hydrolyzed by intracellular esterases to the active Rhod-2 form, which is trapped inside cells. Rhod-2 binds calcium ions, resulting in an increase in fluorescence intensity. The long-wavelength excitation and emission make it valuable for use with laser microscopes.
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| ln Vitro |
1. The working solution for Rhod-2 AM is prepared as follows: 1.1 Prepare the stock solution by dissolving Rhod-2 AM in DMSO to create a 5 mM stock solution. 1.2 Rhod-2 AM working solution preparation: dilute the stock solution to a working solution of 5–10 μM using serum-free cell culture medium or PBS. Please modify the concentration of the Rhod-2 AM working solution to suit the current circumstances. 2. Staining of cells (96-well plate) 2.1 Suspensions a. After centrifuging at 1000g for three to five minutes at 4°C, discard the supernatant. Wash for five minutes each time, twice, using PBS. One x 10^6 cells per milliliter. B. After adding 1 mL of the working solution, let it sit at room temperature for five to thirty minutes. an. After centrifuging at 400 g for three to four minutes at 4°C, discard the supernatant. D. Wash for five minutes each time, twice with PBS. E. In PBS or serum-free cell culture media, resuspend the cells. either flow cytometry monitoring or fluorescence microscopy. Cells that adhere 2.2 a. Adherent cells are cultured on sterile coverslips. B. Lift the coverslip out of the culture media and use the aspirator to remove any extra. an. After adding 100 μL of the working solution and giving the cells a gentle shake to fully cover them, incubate for five to thirty minutes at room temperature. D. Wash for five minutes twice using a medium setting. flow cytometry or fluorescent microscopy.
The in vitro activity of Rhod-2 AM is its function as a fluorescent calcium indicator. The compound is cell-permeable and is hydrolyzed by cytoplasmic esterases to the active Rhod-2 form. Upon binding to calcium ions, the fluorescence intensity increases. Rhod-2 has the longest fluorescent wavelength signal of all the calcium indicators, with excitation/emission maxima of 552/581 nm. It is a fluorescent mitochondrial probe and is used for monitoring intracellular calcium levels. |
| ln Vivo |
Rhod-2 AM is not used as a therapeutic agent. Its in vivo utility is limited to research applications involving the measurement of intracellular calcium levels. The compound is not intended for systemic therapeutic use.
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| Enzyme Assay |
In vitro calcium measurement assays for Rhod-2 AM involve incubating cells with the AM ester in cell culture medium. The compound is taken up by cells, and the AM group is removed by cytoplasmic esterases, trapping the active Rhod-2 inside cells. Cells are then washed and analyzed by fluorescence microscopy or flow cytometry. The fluorescence intensity increases upon binding to calcium ions, providing a measure of intracellular calcium levels.
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| Cell Assay |
For in vitro cellular experiments, Rhod-2 AM is dissolved in DMSO and diluted in cell culture medium. Cells are incubated with the dye for a defined period, washed, and then analyzed by fluorescence microscopy. The red fluorescence (Ex/Em=552/581 nm) provides a measure of mitochondrial calcium levels. The compound is used to study calcium signaling in various cell types.
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| Animal Protocol |
In vivo animal experiments with Rhod-2 AM are not typically performed, as the compound is primarily used as an in vitro probe for calcium measurements in cells.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of Rhod-2 AM have not been characterized, as the compound is a research-use fluorescent probe rather than a drug candidate. The compound has a molecular weight of 1123.94 g/mol and should be stored under appropriate conditions, protected from light.
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| Toxicity/Toxicokinetics |
Rhod-2 AM is intended for research use only and not for human therapeutic or diagnostic applications. Standard laboratory safety precautions should be observed when handling this chemical reagent.
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| References | |
| Additional Infomation |
Rhod-2 dye is an organic bromide salt and thallium dye. It is used as a fluorescent dye. It contains rhod-2(1+).
Rhod-2 AM (CAS#: 145037-81-6) is a cell-permeable acetoxymethyl ester of the red fluorescent calcium indicator Rhod-2. It is used for monitoring intracellular calcium levels, particularly in mitochondria. The compound has no clinical or therapeutic applications. |
| Molecular Formula |
C52H59BRN4O19
|
|---|---|
| Molecular Weight |
1123.94
|
| Exact Mass |
1122.3
|
| CAS # |
145037-81-6
|
| PubChem CID |
25229581
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| Appearance |
Green to dark green ointment
|
| LogP |
2.304
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
23
|
| Rotatable Bond Count |
33
|
| Heavy Atom Count |
76
|
| Complexity |
2060
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CC1=CC(=C(C=C1)N(CC(=O)OCOC(=O)C)CC(=O)OCOC(=O)C)OCCOC2=C(C=CC(=C2)C3=C4C=CC(=CC4=[O+]C5=C3C=CC(=C5)N(C)C)N(C)C)N(CC(=O)OCOC(=O)C)CC(=O)OCOC(=O)C.[Br-]
|
| InChi Key |
RWXWZOWDWYQKBK-UHFFFAOYSA-M
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| InChi Code |
InChI=1S/C52H59N4O19.BrH/c1-32-10-16-42(55(24-48(61)71-28-67-33(2)57)25-49(62)72-29-68-34(3)58)46(20-32)65-18-19-66-47-21-37(11-17-43(47)56(26-50(63)73-30-69-35(4)59)27-51(64)74-31-70-36(5)60)52-40-14-12-38(53(6)7)22-44(40)75-45-23-39(54(8)9)13-15-41(45)52;/h10-17,20-23H,18-19,24-31H2,1-9H3;1H/q+1;/p-1
|
| Chemical Name |
[9-[4-[bis[2-(acetyloxymethoxy)-2-oxoethyl]amino]-3-[2-[2-[bis[2-(acetyloxymethoxy)-2-oxoethyl]amino]-5-methylphenoxy]ethoxy]phenyl]-6-(dimethylamino)xanthen-3-ylidene]-dimethylazanium;bromide
|
| HS Tariff Code |
2934.99.9001
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| 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
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|---|---|
| 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 | 0.8897 mL | 4.4486 mL | 8.8973 mL | |
| 5 mM | 0.1779 mL | 0.8897 mL | 1.7795 mL | |
| 10 mM | 0.0890 mL | 0.4449 mL | 0.8897 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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