Mag-indo-1/AM is a cell-permeable (penetrable) fluorescent indicator of magnesium ions that can also indicate calcium ions (Ex=340-390 nm, Em=410-490 nm). Mag-indo-1/AM may be utilized to measure intracellular magnesium and calcium concentrations.

Physicochemical Properties

Molecular Formula	C33H34N2O17
Molecular Weight	612.58128
Exact Mass	612.196
CAS #	130926-94-2
PubChem CID	2762665
Appearance	Typically exists as solid at room temperature
Density	1.335g/cm3
Boiling Point	789.9ºC at 760mmHg
Flash Point	431.5ºC
Vapour Pressure	6.32E-25mmHg at 25°C
Index of Refraction	1.585
LogP	2.531
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	13
Rotatable Bond Count	21
Heavy Atom Count	44
Complexity	976
Defined Atom Stereocenter Count	0
SMILES	CC(COCOC(CN(C1C=CC(C2=CC3C=CC(=CC=3N2)C(OCOC(=O)C)=O)=CC=1)CC(OCOCC(=O)C)=O)=O)=O
InChi Key	XLQNJLJDRRJLCB-UHFFFAOYSA-N
InChi Code	InChI=1S/C30H32N2O12/c1-19(33)14-39-16-42-28(36)12-32(13-29(37)43-17-40-15-20(2)34)25-8-6-22(7-9-25)26-10-23-4-5-24(11-27(23)31-26)30(38)44-18-41-21(3)35/h4-11,31H,12-18H2,1-3H3
Chemical Name	acetyloxymethyl 2-[4-[bis[2-oxo-2-(2-oxopropoxymethoxy)ethyl]amino]phenyl]-1H-indole-6-carboxylate
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
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro

Guidelines (This is our suggested protocol; it should be adjusted based on your unique requirements as it just offers as a guideline). Intracellular Ca2+ measurement[1]: 1. For 15 minutes at 30°C, incubate a 3 mL cell solution with 10 μM Mag-indo-1/AM. 2. Re-suspend the cells in 5 mL of isolation buffer containing 1 mM CaCl2 after centrifuging them for 1 minute at 500 rpm. 3. To stop dye from leaking out of the cytosol, add 1 mM probenecid to the cell suspension. Then, keep the cells (Mag-indo-1/AM-loaded cells) at room temperature until needed. 4. Centrifuge the pellet, resuspend, and store at room temperature after incubating the Mag-indo-1/AM-loaded cells for two hours at room temperature and for fifteen minutes at 37°C with gentle shaking. 6. A tiny portion of the loaded cells should be placed in an experimental chamber that is set up on an inverted microscope's stage. 7. Determine the Mag-indo-1/AM indicator's fluorescence at Ex=340–390 nm, Em=410–490 nm (the highest emissions of the indicator's Ca2+ bound and Ca2+ free forms). The entire cell's fluorescence is gathered online via a photon counting apparatus.

References

[1]. Griffiths EJ. Calcium handling and cell contraction in rat cardiomyocytes depleted of intracellular magnesium. Cardiovasc Res. 2000 Jul;47(1):116-23. [2]. Measurement of intracellular magnesium concentration in 3T3 fibroblasts with the fluorescent indicator Mag-indo-1. Anal Biochem. 1994 Apr;218(1):170-6.

Solubility Data

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.6324 mL	8.1622 mL	16.3244 mL
	5 mM	0.3265 mL	1.6324 mL	3.2649 mL
	10 mM	0.1632 mL	0.8162 mL	1.6324 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.