EDTA-AM (ethylenediaminetetraacetic acid, acetoxymethyl ester) is a membrane-permeable (penetrable) form of the metal ion chelator EDTA. Living cells are loaded with EDTA-AM by incubating it with EDTA-AM. Once internalized cytosolic esterase decomposes AM, the active ligand EDTA is released, thereby separating the metal ions within the cell. EDTA-AM induces mitotic progression arrest and chromosome decondensation.

Physicochemical Properties

Molecular Formula	C22H32N2O16
Molecular Weight	580.49
Exact Mass	580.175
CAS #	162303-59-5
PubChem CID	91872707
Appearance	Colorless to light yellow liquid
Density	1.3±0.1 g/cm3
Boiling Point	607.0±55.0 °C at 760 mmHg
Flash Point	320.9±31.5 °C
Vapour Pressure	0.0±1.7 mmHg at 25°C
Index of Refraction	1.491
LogP	2.16
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	18
Rotatable Bond Count	27
Heavy Atom Count	40
Complexity	765
Defined Atom Stereocenter Count	0
SMILES	CC(=O)OCOC(=O)CN(CC(OCOC(C)=O)=O)CCN(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O
InChi Key	BYZOHUNEZPRPFB-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H32N2O16/c1-15(25)33-11-37-19(29)7-23(8-20(30)38-12-34-16(2)26)5-6-24(9-21(31)39-13-35-17(3)27)10-22(32)40-14-36-18(4)28/h5-14H2,1-4H3
Chemical Name	acetyloxymethyl 2-[[2-(acetyloxymethoxy)-2-oxoethyl]-[2-[bis[2-(acetyloxymethoxy)-2-oxoethyl]amino]ethyl]amino]acetate
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

Treatment of mitotic cells with EDTA-AM: 1. Gather mitotic cells and seed them onto a 12-well culture plate with coverslips coated in polylysine, 10% FBS added to the DMEM. 2. Once EDTA-AM has been dissolved in 100 mM DMSO, add it to the media that contains 0.02% Pluronic F-127 at a final concentration of 20 μM. Then, incubate the cells for an additional 100 minutes at 37°C. 3. Count the number of interphase and mitotic cells after incubation. After fixing, DAPI staining, and sealing with a portion of p-phenylenediamine (PPDI) solution (20 mM HEPES: pH 7.4, 1 mM MgCl 2, 100 mM KCl, 78% glycerol, 1 mg/mL PPDI), cells on coverslips were prepared.

References

[1]. Watkins CS, et.al. Effects on K+ currents in rat cerebellar granule neurones of a membrane-permeable analogue of the calcium chelator BAPTA. Br J Pharmacol. 1996 Aug;118(7):1772-8. [2]. Maeshima K, et.al. A Transient Rise in Free Mg2+Ions Released from ATP-Mg Hydrolysis Contributes to Mitotic Chromosome Condensation. Curr Biol. 2018 Feb 5;28(3):444-451.e6.

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	1.7227 mL	8.6134 mL	17.2268 mL
	5 mM	0.3445 mL	1.7227 mL	3.4454 mL
	10 mM	0.1723 mL	0.8613 mL	1.7227 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.