Calcein-AM is cell-permeablefluorescent dyeused to determine the cell viability.

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℃. 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].

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

994.86

Solid

C₄₆H₄₆N₂O₂₃

148504-34-1

钙黄绿素-AM

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light

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

DMSO : 100 mg/mL(100.52 mM;Need ultrasonic)

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80℃, 6 months; -20℃, 1 month (protect from light)。-80℃ 储存时，请在 6 个月内使用，-20℃ 储存时，请在 1 个月内使用。

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  Description: Calcein-AM is a fluorescence probe with yellow green fluorescence, it can be used in cell viability assay to differentiate living and dead cells.
  Method: For cell staining.
  1. Cells are plated in 6-well plates and then incubated with indicated compounds at 37℃ and 5% CO₂for 72 h.
  2. Incubate cells with Calcein-AM (5 min).
  3. Use a fluorescence inverted microscope (Olympus IX51, Olympus Corporation, Japan) for image.
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  Description: Calcein-AM is a fluorescent probe with yellow green fluorescence, it can be used in capillary-like tube formation assay.
  Method: For cell staining.
  1. Cells are seeded at a density of 5×10⁴cells/well into Matrigel coated 96-well plates and treated with conditioned medium for 5.5 h under normoxia or hypoxia.
  2. Stain cells with Calcein-AM (0.5 h).
  3. Tubes forming intact networks are captured by Lion Heart (Biotec) and quantified by counting the number of tubes.
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  Description: Calcein-AM is a fluorescent probe with yellow green fluorescence, it can be used in cell proliferation assay.
• 
  Description: Calcein-AM is a fluorescent probe with yellow green fluorescence, it can be used for in vitro capillary tube formation assay.
  Method: For cell staining.
  1. Human umbilical vein endothelial cells (HUVECs) are pre-cultured with other compounds, and the 96-well plate coated with Matrigel is thawed overnight at 4℃ and the Matrigel matrix is allowed to polymerize for 30 min at 37℃.
  2. Trypsin-harvested HUVECs are seeded onto the polymeric Matrigel (1×10⁴cells per well) in 100 μL medium (5 h).
  3. Calcein-AM (100 μL of 2 μM; 40 min) solution is added to per well to stain.
  4. Wash cells for twice and analyze by fluorescent microscopy (Eclipse TE300; Nikon).