Calcein-AM (Calcein acetoxymethyl ester) (Synonyms: Calcein Acetoxymethyl ester, NSC 689290) |
| Catalog No.GC34061 |
Calcein-AM es un colorante vital altamente lipofílico que entra rápidamente en las células viables.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 148504-34-1
Sample solution is provided at 25 µL, 10mM.
Calcein-AM es un colorante vital altamente lipofílico que entra rápidamente en las células viables. Es convertido por esterasas intracelulares en calceína, que produce una señal verde intensa (530 nm) y es retenido por células con membranas plasmáticas intactas. [1]
En experimentos in vitro, se demostró que el ensayo de Calcein-AM se utiliza para evaluar la viabilidad de los eritrocitos humanos después de la incubación (37°C durante 3 y 20 h) en presencia de Ca2+ (2,5 mM) y el ionóforo A 23187 (0,5 μM). [1] 0,05 μM fue la concentración óptima de CAM (Calcein-AM) para teñir las células efectoras, probando con concentraciones de 0,05, 0,1, 0,2 y 0,4 μM. Usando 0,05 μM de CAM para teñir los PBMCs y las células NK expandidas de tres voluntarios normales, los resultados demostraron que no hubo una disminución significativa en la citotoxicidad y que la tinción con CAM no tuvo un efecto significativo en la actividad de las células NK humanas en los PBMCs ni en las células NK expandidas. [2] In vitro, la señal fluorescente de 50μm de Calcein-AM en 1 x 105 linfocitos estaba cerca del nivel de saturación, mientras que la señal emitida por los linfocitos etiquetados con 20μm de Calcein-AM era solo ligeramente menor. [3] Calcein-AM tiene actividad citotóxica contra líneas celulares tumorales humanas (como el linfoma humano U-937-GTB) a bajas concentraciones (2,5 μg/ml). [4] En un experimento in vitro, se demostró que en macrófagos mezclados y células THP-1 (5x105 células/ml), el ensayo de tinción doble Calcein-AM (2 μM)/Ioduro de Propidio (PI) (4,5 μM) se utilizó para cuantificar el número de células vivas y muertas como un ensayo de muerte celular. [5] Además, las células en condrocitos de OA fueron sembradas en placas de 24 pozos (2 × 104 células/pozo), cultivadas durante 4 h, tratadas con 5 μL de Calcein-AM (2 μM) y 5 μL de PI (2 μM) a 37°C en condiciones de ausencia de luz durante 30 min, y luego analizadas bajo un microscopio de fluorescencia. [6]
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.
| Experimento celular [1]: | |
Líneas celulares | EPC cells |
Método de preparación | Cinética de absorción de células EPC cargadas con 5 µM de calceína AM sembradas a una densidad de 1x10^5 células por pozo y cultivadas a 15 °C. La absorción de calceína AM se midió como intensidad de fluorescencia (FI). |
Condiciones de reacción | 5 µM a 15 °C, 1-8 h |
Aplicaciones | La cinética de absorción mostró que, para células EPC sembradas a una densidad de 1x10^5 células por pozo, el marcado con calceína AM aumentó durante las 7 horas probadas. |
Referencias: [1]. Iwanowicz LR, et al. Ensayo citotóxico basado en la liberación de calceína AM para leucocitos de peces. Fish Shellfish Immunol. Feb 2004;16(2):127-37. | |
| 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. | ||
| Prepare stock solution | |||
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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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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
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- Purity: >96.00%
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- SDS (Safety Data Sheet)
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