Calcein-AM 纯度:>96.0% 100μg（2 mg/mL * 50 μL in DMSO）

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Calcein-AM 纯度:>96.0% 100μg（2 mg/mL * 50 μL in DMSO）

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货号：HY-D0041

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Calcein-AM是用于测定细胞活力的可以渗透细胞的荧光染料。

Description

Calcein-AM is cell-permeable fluorescent dye used to determine the cell viability.

In Vitro

The calcein-AM dye used to stain the living cells is shown to have a low spontaneousleakage rate less than 15% in 4 hours at 37°C. Dilutions of targets stained by calcein-AM has a linear relationship with measured fluorescence values. NK cells, LAKs, and CTLs are readily detectable by this microtest. Quantitation of killing and kinetic analysis is readily performed with the test system^[1]. Calcein-AM is pH independent, better retained and more photostable. In addition, the high level of intracellular retention of calcein-AM and its low-level release after incorporation exclude possible cell-monolayer labeling and allow its use in a cell-cell interaction assay. Moreover, the bright fluorescence can easily be detected and measured by a microplate fluorescence reader^[2]. Calcein-AM is a highly lipophilic vital dye that rapidly enters viable cells, is converted by intracellular esterases to calcein that produces an intense green (530-nm) signal, and is retained by cells with intact plasma membrane. From dying or damaged cells with compromised membrane integrity or from cells expressing multidrug resistance protein (MRP), unhydrolyzed substrates and their fluorescent products are rapidly extruded from cells. The calcein-AM assay has been used to assess the cell viability, cytotoxicity and tp quantitate apoptosis^[3].

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].

Solvent & Solubility

In Vitro:

10 mM in 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
10 mM	0.1005 mL	0.5026 mL	1.0052 mL

* Please refer to the solubility information to select the appropriate solvent.

References

[1]. Wang XM, et al. A new microcellular cytotoxicity test based on calcein-AM release. Hum Immunol. 1993 Aug;37(4):264-70.

[2]. Braut-Boucher F, et al. A non-isotopic, highly sensitive, fluorimetric, cell-cell adhesion microplate assay using calceinAM-labeled lymphocytes. J Immunol Methods. 1995 Jan 13;178(1):41-51.

[3]. Bratosin D, et al. Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viabilityand aging. Cytometry A. 2005 Jul;66(1):78-84.

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	[1]. Wang XM, et al. A new microcellular cytotoxicity test based on calcein-AM release. Hum Immunol. 1993 Aug;37(4):264-70. [2]. Braut-Boucher F, et al. A non-isotopic, highly sensitive, fluorimetric, cell-cell adhesion microplate assay using calceinAM-labeled lymphocytes. J Immunol Methods. 1995 Jan 13;178(1):41-51. [3]. Bratosin D, et al. Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viabilityand aging. Cytometry A. 2005 Jul;66(1):78-84.

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

[1]. Wang XM, et al. A new microcellular cytotoxicity test based on calcein-AM release. Hum Immunol. 1993 Aug;37(4):264-70.

[2]. Braut-Boucher F, et al. A non-isotopic, highly sensitive, fluorimetric, cell-cell adhesion microplate assay using calceinAM-labeled lymphocytes. J Immunol Methods. 1995 Jan 13;178(1):41-51.

[3]. Bratosin D, et al. Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viabilityand aging. Cytometry A. 2005 Jul;66(1):78-84.

Molecular Weight	994.86
Formula	C₄₆H₄₆N₂O₂₃
CAS No.	148504-34-1
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
Storage	-20°C, protect from light
Shipping	Room temperature in continental US; may vary elsewhere