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Calcein

Alias: Fluorexon; 1461-15-0; Oftasceine; Oftasceinum; Oftasceina; 2,2',2'',2'''-(((3',6'-Dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-2',7'-diyl)bis(methylene))bis(azanetriyl))tetraacetic acid; Oftasceine [INN];
Cat No.:V6263 Purity: ≥98%
Calcein is a fluorescent dye reagent and self-extinguishing probe that can be used as an indicator of lipid vesicle leakage, and can also be used as a complex indicator for EDTA calcium ion titration and fluorescence determination of calcium ions.
Calcein
Calcein Chemical Structure CAS No.: 1461-15-0
Product category: Fluorescent Dye
This product is for research use only, not for human use. We do not sell to patients.
Size Price Stock Qty
50mg
250mg
Other Sizes

Other Forms of Calcein:

  • Calcein tetraethyl ester
Official Supplier of:
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Top Publications Citing lnvivochem Products
Purity & Quality Control Documentation

Purity: ≥98%

Product Description
Calcein is a fluorescent dye reagent and self-extinguishing probe that can be used as an indicator of lipid vesicle leakage, and can also be used as a complex indicator for EDTA calcium ion titration and fluorescence determination of calcium ions.
Biological Activity I Assay Protocols (From Reference)
Targets
Fluorescent Dye; Emission (Em) = 515; Excitation (Ex) = 489
ln Vitro
In coelomic fluid or ISO-EDTA-incubated coelomocytes, calcein builds up. Calcine fluorescence intensity was higher in coelomic cells cultured in CF as compared to control cultures.
Enzyme Assay
Coelomocytes and hemocytes were stained with the ABC transporter substrate calcein-AM and dye accumulation analyzed under flow cytometry. Reversin 205 (ABCB1 transporter blocker) and MK571 (ABCC1 transporter blocker) were used as pharmacological tools to investigate ABC transporter activity. A different pattern of calcein accumulation was observed in coelomocytes: phagocytes > colorless spherulocytes > vibrate cells > red spherulocytes. The treatment with MK571 increased calcein fluorescence levels in coelomocytes from both species. However, reversin 205 treatment was not able to increase calcein fluorescence in E. esculentus coelomocytes. These data suggest that ABCC1-like transporter activity is present in both sea urchin species, but ABCB1-like transporter activity might only be present in E. lucunter coelomocytes. The activity of ABCC1-like transporter was observed in all cell types from both bivalve species. However, reversin 205 only increased calcein accumulation in hyalinocytes of the oyster C. gasar, suggesting the absence of ABCB1-like transporter activity in all other cell types, including hyalinocytes from the oyster C. gigas. Additionally, our results showed that C. gigas exhibited higher activity of ABCC1-like transporter in all hemocyte types than C. gasar. The present work is the first to characterize ABCB1 and ABCC1-like transporter activity in the immune system cells of sea urchins E. lucunter and E. esculentus and oysters. Our findings encourage the performing studies regarding ABC transporters activity/expression in immune system cells form marine invertebrates under stress conditions and the possible use of ABC transporters as biomarkers[1].
Cell Assay
Examples:
Calcein is a fluorescent probe with yellow green fluorescence, and may be used to determine labile iron pool and measure the Fe(III)-binding affinity with ligands.
Method: For cell staining.
1. Cells (10~6/mL) are loaded by incubation with 20 µM Fe(II) for 2 h at 37°C.
2. Cells are collected by centrifugation (1000 rpm; 5 min) and incubated with Calcein (0.5 µM; 15 min; 37°C) in a D’hanks buffer (pH 7.4).
3. Wash cells for twice by D’hanks buffer (pH 7.4).
4. The fluorescence is monitored by both flow cytometry and a confocal laser scanning microscope.
Description: Calcein is a fluorescent probe with yellow green fluorescence, and may be used in capillary-like tube formation assay.
Method: For cell staining.
1. Cells are seeded at a density of 5×104 cells/well into Matrigel coated 96-well plates and treated with conditioned medium for 5.5 h under normoxia or hypoxia.
2. Stain cells with Calcein (0.5 h).
3. Tubes forming intact networks are captured by Lion Heart (Biotec) and quantified by counting the number of tubes.
Description: Calcein is a fluorescent probe with yellow green fluorescence, and may be used in tube formation assay.
Method: For cell staining.
1. Suspend cultured cells in conditioned medium and seeded into Matrigel coated 96-well plates at a density of 5×104 cells/well for 6 h under normoxia or hypoxia.
2. Stain cells with Calcein (10 min).
3. Tubes forming intact networks are captured by Lion Heart and quantified by the tube length and number of branch points using Image J.
References

[1]. ABCB1 and ABCC1-like transporters in immune system cells from sea urchins Echinometra lucunter and Echinus esculentus and oysters Crassostrea gasar and Crassostrea gigas. Fish Shellfish Immunol. 2017 Sep 5;70:195-203.

Additional Infomation
Calcein is a xanthene dye. It has a role as a fluorochrome. It is functionally related to a fluoran.
Oftasceine is also referred to as Fluorexon. It is used in ophthalmic solutions as a staining agent when fitting soft and hard lenses. It is a fluorescent dye or luminescent agent.
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Exact Mass
622.143
Elemental Analysis
C, 57.88; H, 4.21; N, 4.50; O, 33.41
CAS #
1461-15-0
Related CAS #
Calcein (tetraethyl ester);1170856-93-5
PubChem CID
65079
Appearance
Yellow to orange solid powder
Density
1.7±0.1 g/cm3
Boiling Point
952.7±65.0 °C at 760 mmHg
Melting Point
200 °C
Flash Point
530.0±34.3 °C
Vapour Pressure
0.0±0.3 mmHg at 25°C
Index of Refraction
1.775
LogP
1.56
Hydrogen Bond Donor Count
6
Hydrogen Bond Acceptor Count
15
Rotatable Bond Count
12
Heavy Atom Count
45
Complexity
1070
Defined Atom Stereocenter Count
0
SMILES
O1C(C2=C([H])C([H])=C([H])C([H])=C2C21C1C([H])=C(C([H])([H])N(C([H])([H])C(=O)O[H])C([H])([H])C(=O)O[H])C(=C([H])C=1OC1C([H])=C(C(C([H])([H])N(C([H])([H])C(=O)O[H])C([H])([H])C(=O)O[H])=C([H])C2=1)O[H])O[H])=O
InChi Key
DEGAKNSWVGKMLS-UHFFFAOYSA-N
InChi Code
InChI=1S/C30H26N2O13/c33-21-7-23-19(5-15(21)9-31(11-25(35)36)12-26(37)38)30(18-4-2-1-3-17(18)29(43)45-30)20-6-16(22(34)8-24(20)44-23)10-32(13-27(39)40)14-28(41)42/h1-8,33-34H,9-14H2,(H,35,36)(H,37,38)(H,39,40)(H,41,42)
Chemical Name
2-[[7'-[[bis(carboxymethyl)amino]methyl]-3',6'-dihydroxy-3-oxospiro[2-benzofuran-1,9'-xanthene]-2'-yl]methyl-(carboxymethyl)amino]acetic acid
Synonyms
Fluorexon; 1461-15-0; Oftasceine; Oftasceinum; Oftasceina; 2,2',2'',2'''-(((3',6'-Dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-2',7'-diyl)bis(methylene))bis(azanetriyl))tetraacetic acid; Oftasceine [INN];
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: This product requires protection from light (avoid light exposure) during transportation and storage.
Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vitro)
0.1 M NaOH : ~10 mg/mL (~16.06 mM)
DMSO : ~5 mg/mL (~8.03 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 0.5 mg/mL (0.80 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 5.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

Solubility in Formulation 2: 20 mg/mL (32.13 mM) in 2% NaHCO3 in ddH2O (add these co-solvents sequentially from left to right, and one by one), clear solution; Need ultrasonic and adjust pH to 6 with NaHCO3.

 (Please use freshly prepared in vivo formulations for optimal results.)
Calculator

Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

  • Calculate the Mass of a compound required to prepare a solution of known volume and concentration
  • Calculate the Volume of solution required to dissolve a compound of known mass to a desired concentration
  • Calculate the Concentration of a solution resulting from a known mass of compound in a specific volume
An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
  • Enter 350.26 in the Molecular Weight (MW) box
  • Enter 10 in the Concentration box and choose the correct unit (mM)
  • Enter 5 in the Volume box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
  • Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
  • Enter 25 into the Concentration (End) box and select the correct unit (mM)
  • Enter 25 into the Volume (End) box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box
g/mol

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
Instructions to calculate molar mass (molecular weight) of a chemical compound:
  • To calculate molar mass of a chemical compound, please enter the chemical/molecular formula and click the “Calculate’ button.
Definitions of molecular mass, molecular weight, molar mass and molar weight:
  • Molecular mass (or molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
  • Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.
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Reconstitution Calculator allows you to calculate the volume of solvent required to reconstitute your vial.

  • Enter the mass of the reagent and the desired reconstitution concentration as well as the correct units
  • Click the “Calculate” button
  • The answer appears in the Volume (to add to vial) box
In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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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.

Clinical Trial Information
NCT Number Recruitment interventions Conditions Sponsor/Collaborators Start Date Phases
NCT01892670 COMPLETED Other: Donation of 1 whole blood unit Austere Environment
Blood Bank
Buddy Transfusion
Trauma
Haukeland University Hospital 2013-03 Not Applicable
NCT05948930 RECRUITING Behavioral: Progressive aerobic exercise
Behavioral: Adaptive cognitive training
Behavioral: Combined
Mild Cognitive Impairment University of Maryland, Baltimore 2024-01-08 Not Applicable
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