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  3. Calcein

Calcein is a fluorescent dye and self-quenching probe, used as an indicator of lipid vesicle leakage, and also as a complexometric indicator for titration of calcium ions with EDTA, and for fluorometric determination of calcium.

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Calcein Chemical Structure

Calcein Chemical Structure

CAS No. : 1461-15-0

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Customer Review

Based on 5 publication(s) in Google Scholar

Other Forms of Calcein:

Top Publications Citing Use of Products

    Calcein purchased from MedChemExpress. Usage Cited in: Drug Des Devel Ther. 2022, 16: 3929-3946.

    Calcein and tetracycline labeling of mice femur at 8 weeks. The red arrow indicates the distance between calcein and tetracycline fluorescence labeling.
    • Biological Activity

    • Protocol

    • Purity & Documentation

    • References

    • Customer Review

    Description

    Calcein is a fluorescent dye and self-quenching probe, used as an indicator of lipid vesicle leakage, and also as a complexometric indicator for titration of calcium ions with EDTA, and for fluorometric determination of calcium.

    In Vitro

    Calcein accumulates intracellularly in coelomocytes incubated in coelomic fluid or ISO-EDTA. Coelomocytes incubated in CF show an increase in the calcein fluorescence intensity compared to the control.

    MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

    Molecular Weight

    622.53

    Formula

    C30H26N2O13

    CAS No.
    Appearance

    Solid

    Color

    Pink to red

    Emission (Em)

    515

    Excitation (Ex)

    489

    SMILES

    O=C1OC2(C3=C(OC4=C2C=C(CN(CC(O)=O)CC(O)=O)C(O)=C4)C=C(O)C(CN(CC(O)=O)CC(O)=O)=C3)C5=C1C=CC=C5

    Shipping

    Room temperature in continental US; may vary elsewhere.

    Storage

    4°C, protect from light

    *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

    Solvent & Solubility
    In Vitro: 

    1M NaOH : 50 mg/mL (80.32 mM; Need ultrasonic)

    0.1 M NaOH : 10 mg/mL (16.06 mM; ultrasonic and adjust pH to 11 with NaOH)

    DMSO : 5 mg/mL (8.03 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 1.6063 mL 8.0317 mL 16.0635 mL
    5 mM 0.3213 mL 1.6063 mL 3.2127 mL
    View the Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    • Molarity Calculator

    • Dilution Calculator

    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

    Mass
    =
    Concentration
    ×
    Volume
    ×
    Molecular Weight *

    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

    C1

    ×
    Volume (start)

    V1

    =
    Concentration (final)

    C2

    ×
    Volume (final)

    V2

    In Vivo:

    Select the appropriate dissolution method based on your experimental animal and administration route.

    For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
    To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
    The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

    • Protocol 1

      Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

      Solubility: ≥ 0.5 mg/mL (0.80 mM); Clear solution

      This protocol yields a clear solution of ≥ 0.5 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (5.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

      Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.

    For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
    The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

    • Protocol 1

      Add each solvent one by one:  2% NaHCO3 in ddH2O

      Solubility: 20 mg/mL (32.13 mM); Clear solution; Need ultrasonic and adjust pH to 6 with NaHCO3

    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

    Dosage

    mg/kg

    Animal weight
    (per animal)

    g

    Dosing volume
    (per animal)

    μL

    Number of animals

    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Please enter your animal formula composition:
    %
    DMSO +
    +
    %
    Tween-80 +
    %
    Saline
    Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
    The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
    Calculation results:
    Working solution concentration: mg/mL
    Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

    *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

    The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
    Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
     If the continuous dosing period exceeds half a month, please choose this protocol carefully.
    Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
    Purity & Documentation
    Dyeing Example
    References
    Cell Assay
    [1]

    Coelomocytes suspensions (ISO and CF) are separately incubated with calcein-AM (excitation and emission wavelength: 496 nm and 516 nm, respectively) at a final concentration of 200 nM, during 30 min, at 26°C. Calcein-AM is a nonfluorescent ABC transporter substrate whose intracellular accumulation is inversely proportional to ABC transporter activity. Intracellular esterases convert calcein-AM into the fluorescent dye calcein, which is not an ABC transporter substrate, thereby accumulating the dye inside the cell. Therefore, a high fluorescence signal indicates low ABC transporter activity whereas a low fluorescence signal indicates high activity. The fluorescence of samples is measured by flow cytometer. The experiment is repeated six times in duplicates.

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    References

    Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
    DMSO / 0.1 M NaOH / 1M NaOH 1 mM 1.6063 mL 8.0317 mL 16.0635 mL 40.1587 mL
    5 mM 0.3213 mL 1.6063 mL 3.2127 mL 8.0317 mL
    0.1 M NaOH / 1M NaOH 10 mM 0.1606 mL 0.8032 mL 1.6063 mL 4.0159 mL
    15 mM 0.1071 mL 0.5354 mL 1.0709 mL 2.6772 mL
    1M NaOH 20 mM 0.0803 mL 0.4016 mL 0.8032 mL 2.0079 mL
    25 mM 0.0643 mL 0.3213 mL 0.6425 mL 1.6063 mL
    30 mM 0.0535 mL 0.2677 mL 0.5354 mL 1.3386 mL
    40 mM 0.0402 mL 0.2008 mL 0.4016 mL 1.0040 mL
    50 mM 0.0321 mL 0.1606 mL 0.3213 mL 0.8032 mL
    60 mM 0.0268 mL 0.1339 mL 0.2677 mL 0.6693 mL
    80 mM 0.0201 mL 0.1004 mL 0.2008 mL 0.5020 mL
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    Calcein Related Classifications

    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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