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Calcein-AM (Calcein acetoxymethyl ester) (Synonyms: Calcein Acetoxymethyl ester, NSC 689290)

Catalog No.GC34061

Calcein-AM (éster acetoximetil de calceína) es un tinte fluorescente permeable a las células utilizado para determinar la viabilidad celular.

Products are for research use only. Not for human use. We do not sell to patients.

Calcein-AM (Calcein acetoxymethyl ester) Chemical Structure

Cas No.: 148504-34-1

Tamaño Precio Disponibilidad Cantidad
500μg
149,00 $
Disponible
1mg
282,00 $
Disponible

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Sample solution is provided at 25 µL, 10mM.

Product has been cited by 5 publications

Protocol

Cell experiment [1]:

Cell lines

EPC cells

Preparation Method

Uptake kinetics of EPC cells loaded with 5 µM calcein AM seeded at 1x105 cells well-1 and cultured at 15 °C. Calcein AM uptake was measured as fluorescence intensity (FI).

Reaction Conditions

5 µM at 15 °C, 1-8h

Applications

Uptake kinetics showed that for EPC cells seeded at a density of 1x105 cells well-1 calcein AM labelling increased throughout the 7 h tested.

References:

[1]. Iwanowicz LR, et al. Calcein AM release-based cytotoxic cell assay for fish leucocytes. Fish Shellfish Immunol. 2004 Feb;16(2):127-37.

Background

Calcein-AM es un colorante vital altamente lipofílico que entra rápidamente en las células viables. Es convertido por esterasas intracelulares a calceína, la cual produce una señal verde intensa (530 nm) y es retenida por células con membrana plasmática intacta.[1]

El experimento in vitro demostró que el ensayo de calceína-AM se utilizó para evaluar la viabilidad de los glóbulos rojos humanos después de la incubación (a 37°C durante 3 y 20 horas) en presencia de Ca2+ (2,5 mM) e ionóforo A23187 (0,5 μM). La concentración óptima de CAM (calceína-AM) para teñir las células efectoras fue de 0,05 μM al probar 0,05, 0,1, 0,2 y 0.4 μM. Al utilizar una concentración de CAM del 0.05μm para teñir PBMCs y células NK expandidas a partir de tres voluntarios normales, los resultados demostraron que no hubo disminución significativa en la citotoxicidad y que el tinte CAM no tuvo un efecto significativo sobre la actividad celular NK humana en PBMCs o células NK expandidas. En vitro ,la señal fluorescente del AM-calceína a una concentración de50μm era cercana al nivel máximo mientras que la señal emitida por linfocitos marcados con AM-calceína a una concentraciónde20μm era solo ligeramente menor.Calceina-AM tiene actividad citotóxica contra líneas celulares tumorales humanas como el linfoma humano U-937-GTB a bajas concentraciones(2.5ug/ml). El experimento in vitro demostró que en macrófagos mixtos y células THP-1(5x105células/ml), se utilizó un ensayo doble Calcein-AM/PI para cuantificar el número de células vivas y muertas como un ensayo de muerte celular. Además, las células en condrocitos OA se sembraron en placas de 24 pocillos (2 × 104 células/pocillo), se cultivaron durante 4 h, las células fueron tratadas con 5 μL Calcein-AM (2 μM) y 5 μL PI (2 μM) a37°C en condiciones sin luz durante30 min,y luego se analizaron bajo un microscopio de fluorescencia.

References:
[1].Bratosin D, et al. Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viability and aging. Cytometry A. 2005 Jul;66(1):78-84.
[2].Jang YY, et al. An improved flow cytometry-based natural killer cytotoxicity assay involving calcein AM staining of effector cells. Ann Clin Lab Sci. 2012 Winter;42(1):42-9.
[3].Braut-Boucher F, et al. A non-isotopic, highly sensitive, fluorimetric, cell-cell adhesion microplate assay using calcein AM-labeled lymphocytes. J Immunol Methods. 1995 Jan 13;178(1):41-51.
[4].Liminga G, et al. Cytotoxic effect of calcein acetoxymethyl ester on human tumor cell lines: drug delivery by intracellular trapping. Anticancer Drugs. 1995 Aug;6(4):578-85.
[5].Xiang N, et al. Gardnerella vaginalis induces NLRP3 inflammasome-mediated pyroptosis in macrophages and THP-1 monocytes. Exp Ther Med. 2021 Oct;22(4):1174.
[6].Zhang L, et al. MicroRNA-140-5p represses chondrocyte pyroptosis and relieves cartilage injury in osteoarthritis by inhibiting cathepsin B/Nod-like receptor protein 3. Bioengineered. 2021 Dec;12(2):9949-9964.

Chemical Properties

Cas No. 148504-34-1 SDF
Sinónimos Calcein Acetoxymethyl ester, NSC 689290
Chemical Name tetrakis(acetoxymethyl) 2,2',2'',2'''-(((3',6'-diacetoxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-4',5'-diyl)bis(methylene))bis(azanetriyl))tetraacetate
Canonical 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
Formula C46H46N2O23 M.Wt 994.86
Solubility 10 mg/mL in EtOH, MeOH, DMSO, DMF with gentle heating (37°C) for 10 minutes. Storage -20°C, protect from light
General tips Please select the appropriate solvent to prepare the stock solution according to the solubility of the product in different solvents; once the solution is prepared, please store it in separate packages to avoid product failure caused by repeated freezing and thawing.Storage method and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored at -20°C, please use it within 1 month.
To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time.
Shipping Condition Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request.

Complete Stock Solution Preparation Table

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1 mg 5 mg 10 mg
1 mM 1.0052 mL 5.0258 mL 10.0517 mL
5 mM 0.201 mL 1.0052 mL 2.0103 mL
10 mM 0.1005 mL 0.5026 mL 1.0052 mL
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Review for Calcein-AM (Calcein acetoxymethyl ester)

Average Rating: 5 ★★★★★ (Based on Reviews and 1 reference(s) in Google Scholar.)

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