Calcium Green-5N AM 

Calcium Green-5N AM is a Ca²⁺-sensitive fluorescent probe (K_d = 14 μM). Calcium Green-5N AM shows enhanced fluorescence intensity upon binding to Ca²⁺, and it can monitor free Ca²⁺ in neurons^[1][2][3].

Guide (The following is our recommended solution. This solution is merely a guideline and should be modified according to your specific needs.)
1.\tIsolate Mitochondria: Prepare mitochondria from tissues (e.g., heart, liver) using differential centrifugation.
2.\tSetup Assay: Add isolated mitochondria (0.1-0.5 mg/mL) to a 96-well plate or spectrofluorometer cuvette containing respiratory buffer.
3.\tAdd Dye: Add Calcium Green-5N to a final concentration of 1 μM.
4.\tEnergize: Add substrate (e.g., 5 mM glutamate + 5 mM malate) and allow 2-5 min to incubate at room temperature, protecting from light.
5.\tInject Calcium: Program a plate reader to inject Cacl₂ pulses (e.g., 5-10 μM per injection) every 1-2 minutes.
6.\tAnalyze: Monitor the increase in fluorescence. A sudden, sharp increase indicates the mitochondria are overloaded, the MPTP has opened, and is being released.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Calcium Green-5N AM (4 μM) labels high (≥ 1 μM) Ca²⁺ levels in the acrosomal crescent of capacitated mouse sperm, and this signal is lost upon thapsigargin-induced acrosomal exocytosis^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

1335.96

C₅₉H₅₂Cl₂N₄O₂₈^2-

216699-33-1

Oil

Brown to reddish brown

521

492

O=C1OC2(C3=CC(Cl)=C(C=C3OC4=CC(OC(C)=O)=C(C=C42)Cl)OC(C)=O)C5=CC=C(C(NC6=CC=C(N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)C(OCCOC7=CC([N-]([O-])=O)=CC=C7N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)=C6)=O)C=C15

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light, stored under nitrogen

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

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Herrick SB, et al. The acrosomal vesicle of mouse sperm is a calcium store. J Cell Physiol. 2005;202(3):663-671.  [Content Brief]

  [2]. Thomas D, et al. A comparison of fluorescent Ca2+ indicator properties and their use in measuring elementary and global Ca2+ signals. Cell Calcium. 2000;28(4):213-223.  [Content Brief]

  [3]. Rajdev S, et al. Calcium green-5N, a novel fluorescent probe for monitoring high intracellular free Ca2+ concentrations associated with glutamate excitotoxicity in cultured rat brain neurons. Neurosci Lett. 1993;162(1-2):149-152.  [Content Brief]