Calcein is afluorescent dyeand self-quenching probe, used as an indicator of lipid vesicle leakage, and also as a complexometric indicator for titration of calcium ions with EDTA, and for fluorometric determination of calcium.

Calcein accumulates intracellularly in coelomocytes incubated in coelomic fluid or ISO-EDTA. Coelomocytes incubated in CF show an increase in the calcein fluorescence intensity compared to the control.

622.53

Solid

C₃₀H₂₆N₂O₁₃

1461-15-0

钙黄绿素

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

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

DMSO : ≥ 100 mg/mL(160.63 mM)

0.1 M NaOH : 10 mg/mL(16.06 mM;ultrasonic and adjust pH to 11 with NaOH)

*"≥" means soluble, but saturation unknown.

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。

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照In Vitro方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 2% NaHCO3 in ddH2O

  Solubility: 20 mg/mL (32.13 mM); Clear solution; Need ultrasonic and adjust pH to 6 with NaHCO3

• 
  Description: Calcein is a fluorescent probe with yellow green fluorescence, it can be used to determine labile iron pool and measure the Fe(III)-binding affinity with ligands.
  Method: For cell staining.
  1. Cells (10⁶/mL) are loaded by incubation with 20 μM Fe(II) for 2 h at 37℃.
  2. Cells are collected by centrifugation (1000 rpm; 5 min) and incubated with Calcein (0.5 μM; 15 min; 37℃) in a D’hanks buffer (pH 7.4).
  3. Wash cells for twice by D’hanks buffer (pH 7.4).
  4. The fluorescence is monitored by both flow cytometry (BD FACS Caliber) and a confocal laser scanning microscope (Leica TCS SP).
• 
  Description: Calcein 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 (0.5 h).
  3. Tubes forming intact networks are captured by Lion Heart (Biotec) and quantified by counting the number of tubes.
• 
  Description: Calcein is a fluorescent probe with yellow green fluorescence, it can be used in tube formation assay.
  Method: For cell staining.
  1. Suspend cultured cells in conditioned medium and seeded into Matrigel coated 96-well plates at a density of 5×10⁴cells/well for 6 h under normoxia or hypoxia.
  2. Stain cells with Calcein (10 min).
  3. Tubes forming intact networks are captured by Lion Heart (Biotek) and quantified by the tube length and number of branch points using Image J.