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  4. Calcein-AM

Calcein-AM  (Synonyms: Calcein acetoxymethyl ester)

Cat. No.: HY-D0041 Purity: 99.08%
SDS COA Handling Instructions

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Calcein AM, has cell membrane permeability and can easily enter the cell. Calcein AM has no fluorescence and is hydrolyzed by endogenous esterase in the cell to produce polar molecule Calcein (Calcein), which has strong negative charge and cannot permeate the cell membrane. Calcein can emit strong green fluorescence, so it is often used with Propidium Iodide for cell viability/virulence detection, excitation/emission wavelength: 494/515 nm.

For research use only. We do not sell to patients.

Calcein-AM Chemical Structure

Calcein-AM Chemical Structure

CAS No. : 148504-34-1

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1 mg USD 276 In-stock
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Customer Review

Based on 56 publication(s) in Google Scholar

Calcein-AM Related Antibodies

Top Publications Citing Use of Products

49 Publications Citing Use of MCE Calcein-AM

IF

    Calcein-AM purchased from MedChemExpress. Usage Cited in: Bioact Mater. 2022 Aug 11;21:20-31.  [Abstract]

    HUVECs proliferation is assessed by Calcein-AM, and the results show that HP-β-CD and HP-β-CD@Res could effectively promote the proliferation of HUVECs.

    Calcein-AM purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2022 Oct;9(30):e2203031.  [Abstract]

    XPS analysis of CaALN powder before and after immersing in FeCl3 solution for 24 h. Detection of intracellular Fe concentration using Calcein-AM in MGC 803 cells after treatment with NaALN, CaALN, and DFX at 25 mg/L for 24 h by FCM analysis. Then these cells are washed with PBS and incubated with Calcein-AM (0.5 μM) for 15 min at 37 °C in the dark.

    Calcein-AM purchased from MedChemExpress. Usage Cited in: Pharmaceuticals. 2022 Aug 26;15(9):1059.

    For U251 cells, Calcein-AM (10 μM) solution and 0.5 μL PI staining solution (15 μM) is added into each well, stained for 10 min.

    Calcein-AM purchased from MedChemExpress. Usage Cited in: Antioxidants (Basel). 2021 May 18;10(5):798.  [Abstract]

    HeLa cells are pretreated with 20 μM Fe(II) for 2 h and then incubated with Calcein-AM ( 0.5 μM; for 15 min at 37 °C). After 15 min, cells are washed and incubated with or without CAPE and its analogues for 15 min. The fluorescence intensity of cells is measured by flow cytometry.

    Calcein-AM purchased from MedChemExpress. Usage Cited in: Biochem Pharmacol. 2021 Jun;188:114583.  [Abstract]

    Cells are incubated with fluorescence probes Calcein-AM (1 μM) and propidium iodide (5 μM) for 5 min and imaged through fluorescence inverted microscope.

    Calcein-AM purchased from MedChemExpress. Usage Cited in: Acta Pharm Sin B. 2020 Sep;10(9):1694-1708.  [Abstract]

    HUVECs are pre-treated with SMI or Rd for 24 h and seeded on Matrigel for 5.5 h followed by 0.5 h of Calcein-AM staining.

    Calcein-AM purchased from MedChemExpress. Usage Cited in: EMBO Rep. 2019 Jul;20(7):e47563.  [Abstract]

    Cells are trypsinized, washed twice with 0.5 ml PBS, and then incubated with 0.2 μM Calcein‐AM at 37°C for 15 min. Then, the cells were washed twice with 0.5 ml PBS and either incubated with 100 μM Deferiprone or left untreated at 37°C for 1 h. The cells are analyzed with a flow cytometer. Calcein-AM is excited at 488 nm, and fluorescence is measured at 525 nm.

    • Biological Activity

    • Protocol

    • Purity & Documentation

    • References

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    Description

    Calcein AM, has cell membrane permeability and can easily enter the cell. Calcein AM has no fluorescence and is hydrolyzed by endogenous esterase in the cell to produce polar molecule Calcein (Calcein), which has strong negative charge and cannot permeate the cell membrane. Calcein can emit strong green fluorescence, so it is often used with Propidium Iodide for cell viability/virulence detection, excitation/emission wavelength: 494/515 nm[1].

    In Vitro

    General Protocol

    1.Preparation of Calcein AM working solution
    Preparation of 5 mM stock solution with DMSO.
    Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of working solution.
    Note: Please adjust the concentration of Calcein AM working solution according to the actual situation.

    2.Cell staining
    2.1 Suspension cells（6-well plate）
    a.Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×106/mL.
    b.Add 1 mL of working solution, and then incubate at room temperature for 30-45 minutes.
    c.Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
    d.Wash twice with PBS, 5 minutes each time.
    e.Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or fluorescence microplate readers.
    2.2 Adherent cells
    a.Culture adherent cells on sterile coverslips.
    b.Remove the coverslip from the medium and aspirate excess medium.
    c.Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 30-45 minutes.
    d.Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or fluorescence microplate readers.

    Storage
    -20°C
    Protect from light

    Precautions
    1.Please adjust the concentration of Calcein AM working solution according to the actual situation.
    2.This product is for R&D use only, not for drug, household, or other uses.
    3.For your safety and health, please wear a lab coat and disposable gloves to operate.

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

    Calcein-AM Related Antibodies
    In Vivo

    Calcein-AM is found to be suitable for in vivo studies, because it has no deleterious effects on cell function and is, indeed, a marker of cell viability[2].

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

    994.86

    Formula

    C46H46N2O23
    CAS No.
    Appearance

    Solid

    Color

    White to off-white

    Emission (Em)

    515
    Excitation (Ex)

    485
    SMILES

    O=C1OC2(C(C=C(CN(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)C(OC(C)=O)=C3)=C3OC4=CC(OC(C)=O)=C(CN(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)C=C24)C5=C1C=CC=C5
    Shipping

    Room temperature in continental US; may vary elsewhere.
    Storage

    -20°C, protect from light

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

    Solvent & Solubility
    In Vitro: 

    DMSO : 100 mg/mL (100.52 mM; Need ultrasonic; 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 1.0052 mL 5.0258 mL 10.0517 mL
    5 mM 0.2010 mL 1.0052 mL 2.0103 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 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.
    Calculation results:
    Working solution concentration: mg/mL
    Purity & Documentation

    Purity: 99.08%

    SDS (393 KB)

    COA (254 KB) HNMR (332 KB) RP-HPLC (269 KB) LCMS (91 KB)

    Handling Instructions (2659 KB)
    Dyeing Example
    References
    Cell Assay
    [1][2][3]

    K562, Daudi, and Chang liver cells are labeled with calcein-AM. Calcein-AM's excitation and emission wavelengths are 496 nm and 520 nm, respectively. The filter/mirror combination used to detect calcein-AM's green fluorescence includes the 490-nm excitation and 520-nm emission filters with a dichroic mirror. Differences in the automatic fluorescence readings between the test and control wells determine the results[1]. A simple and sensitive cell-cell adhesion microplate assay is established using the calcein-AM. The procedure involves three steps: the labeling of lymphocytes with an adequate concentration of calcein-AM (20 μM) during a short incubation period (30 min); the adhesion of 2×105 labeled lymphocytes per well to confluent keratinocyte or fibroblast monolayers grown in microtiter plates for 90 min; and, finally, measurement of the fluorescent signal utilizing a new system of cold-light microfluorimetry[2]. Cells are incubated for 15 min in 1 mL of a 1% saponin solution in PBS buffer, pH 7.4, containing 0.05% sodium azide. After saponin permeabilization, 4×105 RBCs in suspension in PBS buffer containing 0.1% saponin and 0.05% sodium azide are incubated (37°C in the dark for 45 min) with calcein-AM to a final concentration of 5 μM, ished three times with the same PBS buffer containing 0.1% saponin and 0.05% sodium azide, and the cell viability is analyzed by flow cytometry[3].

    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 1 mM 1.0052 mL 5.0258 mL 10.0517 mL 25.1292 mL
    5 mM 0.2010 mL 1.0052 mL 2.0103 mL 5.0258 mL
    10 mM 0.1005 mL 0.5026 mL 1.0052 mL 2.5129 mL
    15 mM 0.0670 mL 0.3351 mL 0.6701 mL 1.6753 mL
    20 mM 0.0503 mL 0.2513 mL 0.5026 mL 1.2565 mL
    25 mM 0.0402 mL 0.2010 mL 0.4021 mL 1.0052 mL
    30 mM 0.0335 mL 0.1675 mL 0.3351 mL 0.8376 mL
    40 mM 0.0251 mL 0.1256 mL 0.2513 mL 0.6282 mL
    50 mM 0.0201 mL 0.1005 mL 0.2010 mL 0.5026 mL
    60 mM 0.0168 mL 0.0838 mL 0.1675 mL 0.4188 mL
    80 mM 0.0126 mL 0.0628 mL 0.1256 mL 0.3141 mL
    100 mM 0.0101 mL 0.0503 mL 0.1005 mL 0.2513 mL
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    Calcein-AM 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.

    Keywords:

    Calcein-AM148504-34-1Calcein acetoxymethyl esterFluorescent DyeInhibitorinhibitorinhibit

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