Fluo-4 AM
Based on 63 publication(s) in Google Scholar
Fluo-4 AM is a cell-permeable Ca2+ (calcium ion) indicator (Ex/Em = 485/526 nm).
For research use only. We do not sell to patients.
- Purity: 99.84%
- CAS No.: 273221-67-3
- Formula: C51H50F2N2O23
- Molecular Weight:1096.94
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Storage:
-20°C, sealed storage, away from moisture and light
* The compound is unstable in solutions, freshly prepared is recommended.
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Protocol
- Purity & Documentation
- References
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Help & FAQs
Publications Citing Use of MedChemExpress (MCE) Fluo-4 AM
More- Adv Mater. 2025 Sep 7:e08101. [Abstract]
- Bioact Mater. 2022 May 21;20:81-92. [Abstract]
- Nat Aging. 2026 Feb;6(2):349-367. [Abstract]
- ACS Nano. 2025 Dec 23;19(50):42022-42045. [Abstract]
- ACS Nano. 2025 Sep 30;19(38):33828-33840. [Abstract]
- Nat Commun. 2025 Dec 16;17(1):272. [Abstract]
- Adv Sci (Weinh). 2026 Mar 12:e22572. [Abstract]
- Chem Eng J. 2024 Dec 15.
- J Adv Res. 2024 Aug:62:187-198. [Abstract]
- J Nanobiotechnology. 2025 Aug 26;23(1):590. [Abstract]
- J Immunother Cancer. 2025 Oct 5;13(10):e012182. [Abstract]
- J Exp Med. 2025 Dec 1;222(12):e20250153. [Abstract]
- Mater Today Bio. 2026 May 29:38:103285. [Abstract]
- Mater Today Bio. 2025 Nov 7:35:102527. [Abstract]
- ISME J. 2023 Dec;17(12):2426-2440. [Abstract]
- Ultrason Sonochem. 2024 Feb:103:106768. [Abstract]
- Cell Death Dis. 2025 Jan 26;16(1):43. [Abstract]
- Acta Biomater. 2023 Jul 1:164:626-640. [Abstract]
- Nano Res. 2026 Mar 31.
- Nano Res. 2022;15(10):9286-9297. [Abstract]
- Int J Biol Macromol. 2025 Dec;334(Pt 2):149080. [Abstract]
- Phytomedicine. 2025 Dec:149:157553. [Abstract]
- Free Radic Biol Med. 2026 Aug 16:252:214-226. [Abstract]
- Free Radic Biol Med. 2026 Jun 7:254:307-322. [Abstract]
- Brain Behav Immun. 2025 Jul:127:341-357. [Abstract]
- J Transl Med. 2024 Jun 6;22(1):538. [Abstract]
- J Transl Med. 2023 Oct 10;21(1):711. [Abstract]
- Cell Death Discov. 2021 Feb 10;7(1):31. [Abstract]
- Cell Rep. 2025 Nov 25;44(11):116543. [Abstract]
- Phytother Res. 2024 Apr;38(4):1990-2006. [Abstract]
- Neurosci Bull. 2025 May 29. [Abstract]
- Chin Med. 2025 Oct 6;20(1):162. [Abstract]
- Ocul Surf. 2025 Jan 18:36:106-118. [Abstract]
- Biochem Pharmacol. 2024 Feb:220:115962. [Abstract]
- NanoImpact. 2025 Jun 15:39:100572. [Abstract]
- J Ethnopharmacol. 2026 Feb 28:357:120906. [Abstract]
- Inflammation. 2026 Jan 30;49(1):72. [Abstract]
- aBIOTECH. 2023 Sep 20;4(3):185-201. [Abstract]
- Int J Mol Sci. 2026 Jan;27(13):5641.
- Front Pharmacol. 2025 Nov 18:16:1697815. [Abstract]
- Front Pharmacol. 2022 May 17;13:855626. [Abstract]
- Int Immunopharmacol. 2025 May 30:160:114918. [Abstract]
- Eur J Pharmacol. 2023 May 5:946:175647. [Abstract]
- Toxicology. 2025 Jun 4:154212. [Abstract]
- ACS Omega. 2025 Aug 21;10(34):39192-39202. [Abstract]
- Microorganisms. 2026 Mar 28;14(4):771. [Abstract]
- iScience. 2021 Oct 12;24(11):103261. [Abstract]
- Cell Calcium. 2026 Mar:134:103122. [Abstract]
- J Dermatol Sci. 2025 Apr;118(1):29-37. [Abstract]
- Exp Cell Res. 2025 Apr 1;447(1):114493. [Abstract]
- Int J Cardiol. 2021 Sep 1:338:185-195. [Abstract]
- Mycotoxin Res. 2025 Dec 27;42(1):15. [Abstract]
- J Dent Sci. 2023 Oct;18(4):1685-1692. [Abstract]
- Mol Biol Rep. 2023 Oct;50(10):8237-8247. [Abstract]
- J Dig Dis. 2025 Apr 14. [Abstract]
- Biophys Chem. 2024 Dec 3:318:107367. [Abstract]
- SSRN. 2026 May 27.
- Res Sq. 2026 Apr 9.
- bioRxiv. 2025 Dec 22.
- bioRxiv. 2025 Jul 18.
- Research Square Preprint. 2024 Feb 16.
- Evid Based Complement Alternat Med. 2021 Jul 16;2021:6622848. [Abstract]
- Oxid Med Cell Longev. 2021 Feb 10;2021:8884922. [Abstract]
Customer Validation & Images
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IF
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Biological Activity
Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)
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1. Solution preparation[1]
1.1 Preparation of storage solution
Solvent: DMSO
Concentration: 1 mM is recommended (e.g. weigh 100 μg Fluo-4 AM and dissolve in 96.16 μL solvent).
Storage: Use as soon as possible and avoid long-term storage.
1.2 Preparation of working solution
Dilute with buffer (e.g. HHBS containing 0.04% Poloxamer 407 (F127) (HY-D1005)) to 1 M (optimized according to the experiment).
Note: The working solution should be prepared and used immediately, and protected from light.
2. Cell staining
2.1 Count the cells and take 106 cells from each sample (control and experiment/s).
2.2 Collect the cells (5 min, 3000×g, 4 °C) and wash once in PBS.
2.3 Resuspend the cells in PBS and add Fluo-4-AM (1 mM stock) to a final concentration of 1 μM. Incubate at 37 °C for 1 h.
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4 Wash the cells three times (2 min, 3000×g) with PBS and finally resuspend in 1 mL PBS.
2.5 Evaluate the staining by flow cytometry (Ex/Em = 485/526 nm).
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
526
485
Chemical Information
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CAS No. 273221-67-3
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Appearance Solid
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Molecular Weight 1096.94
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Formula C51H50F2N2O23
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Color Brown to red
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SMILES
O=C(OCOC(C)=O)CN(C1=CC=C(C2=C3C=C(F)C(C=C3OC4=C2C=C(F)C(OCOC(C)=O)=C4)=O)C=C1OCCOC5=CC(C)=CC=C5N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
-20°C, sealed storage, away from moisture and light
* The compound is unstable in solutions, freshly prepared is recommended.
Publications (63)
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Journal Impact Factor
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Most Recent
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Adv Mater
Ultrasound-Activated Piezoelectric Nanoparticles Targeting and Activating NK Cells for Tumor Immunotherapy. [Abstract]2025 Sep 7:e08101. PMID: 40916189 -
Bioact Mater
Electrical charge on ferroelectric nanocomposite membranes enhances SHED neural differentiation. [Abstract]2022 May 21;20:81-92. PMID: 35633875 -
Nat Aging
Senotoxins target senescence via lipid binding specificity, ion imbalance and lipidome remodeling. [Abstract]2026 Feb;6(2):349-367. PMID: 41526742 -
ACS Nano
Osteogenic Differentiation Triggered by Intracellular Magnetoelectric Stimulation of Core-Shell Nanotransducers under Remotely Applied Magnetic Fields. [Abstract]2025 Dec 23;19(50):42022-42045. PMID: 41347717 -
ACS Nano
Exploring Metabolic Abnormalities of Amino Acids in Preeclampsia-Associated Cardiac Physiological Dysfunction via Cardiomyocyte-Based Multimodal Biosensing. [Abstract]2025 Sep 30;19(38):33828-33840. PMID: 40939054 -
Nat Commun
Polypeptide sono-adjuvant for ultrasound-activatable regulation of innate immunity and cancer vaccination therapy. [Abstract]2025 Dec 16;17(1):272. PMID: 41402324 -
Adv Sci (Weinh)
2026 Mar 12:e22572. PMID: 41816968 -
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J Adv Res
Calcium Influx: An Essential Process by which α-Synuclein Regulates Morphology of Erythrocytes. [Abstract]2024 Aug:62:187-198. PMID: 37714326 -
J Nanobiotechnology
Sparking angiogenesis by carbon monoxide-rich gold nanoparticles obtained by pulsed laser driven CO2 reduction reaction. [Abstract]2025 Aug 26;23(1):590. PMID: 40859339 -
J Immunother Cancer
Macrophage NLRP3 activation and IL-1β release drive osimertinib-induced antitumor immunity. [Abstract]2025 Oct 5;13(10):e012182. PMID: 41052880 -
J Exp Med
Heterozygous human JAK2V617F activates AhR to drive essential thrombocythemia and promote thrombosis. [Abstract]2025 Dec 1;222(12):e20250153. PMID: 41091148 -
Mater Today Bio
Magnetic hydrogel-mediated periodic stimulation enhances mesenchymal stem cell small extracellular vesicle release. [Abstract]2026 May 29:38:103285. PMID: 42256076 -
Mater Today Bio
A bufalin and CRISPR/Cas9 ribonucleoprotein-loaded calcium lactate nanomedicine for pyroptosis/apoptosis and synergistic cancer immunotherapy. [Abstract]2025 Nov 7:35:102527. PMID: 41322154 -
ISME J
Ginsenoside Rg3 enriches SCFA-producing commensal bacteria to confer protection against enteric viral infection via the cGAS-STING-type I IFN axis. [Abstract]2023 Dec;17(12):2426-2440. PMID: 37950067 -
Ultrason Sonochem
Enhanced macromolecular substance extravasation through the blood-brain barrier via acoustic bubble-cell interactions. [Abstract]2024 Feb:103:106768. PMID: 38241945 -
Cell Death Dis
MAPK4 inhibits the early aberrant activation of B cells in rheumatoid arthritis by promoting the IRF4-SHIP1 signaling pathway. [Abstract]2025 Jan 26;16(1):43. PMID: 39863600 -
Acta Biomater
Hydroxyapatite nanoparticles promote TLR4 agonist-mediated anti-tumor immunity through synergically enhanced macrophage polarization. [Abstract]2023 Jul 1:164:626-640. PMID: 37086827 -
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Nano Res
Hydroxyapatite nanoparticles drive the potency of Toll-like receptor 9 agonist for amplified innate and adaptive immune response. [Abstract]2022;15(10):9286-9297. PMID: 35911480 -
Int J Biol Macromol
Mimotopes of calcitonin gene-related peptide (CGRP) screened from Fv-antibody library: antagonists to CGRP receptor. [Abstract]2025 Dec;334(Pt 2):149080. PMID: 41253226 -
Phytomedicine
Liquiritigenin attenuates thrombosis by inhibiting P2Y12R-mediated Src/PLCγ2 signaling and cytoplasmic Ca2+ mobilization in platelets. [Abstract]2025 Dec:149:157553. PMID: 41273866 -
Free Radic Biol Med
TSPO integrates redox and immune homeostasis in dermal fibroblasts to restrain vitiligo pathogenesis. [Abstract]2026 Aug 16:252:214-226. PMID: 42097319 -
Free Radic Biol Med
PARS2 deficiency impairs mitochondrial homeostasis and activates ferroptotic to drive developmental and epileptic encephalopathy. [Abstract]2026 Jun 7:254:307-322. PMID: 42259431 -
Brain Behav Immun
Mechanistic insights into the neuroprotective effects of low-intensity transcranial ultrasound stimulation in post-cardiac arrest brain injury: Modulation of the Piezo1-Dkk3/PI3K-Akt pathway. [Abstract]2025 Jul:127:341-357. PMID: 40118226 -
J Transl Med
CAPN2 promotes apalutamide resistance in metastatic hormone-sensitive prostate cancer by activating protective autophagy. [Abstract]2024 Jun 6;22(1):538. PMID: 38844946 -
J Transl Med
Matrix stiffness induces Drp1-mediated mitochondrial fission through Piezo1 mechanotransduction in human intervertebral disc degeneration. [Abstract]2023 Oct 10;21(1):711. PMID: 37817199 -
Cell Death Discov
IP3R1 regulates Ca2+ transport and pyroptosis through the NLRP3/Caspase-1 pathway in myocardial ischemia/reperfusion injury. [Abstract]2021 Feb 10;7(1):31. PMID: 33568649 -
Cell Rep
2025 Nov 25;44(11):116543. PMID: 41206865 -
Phytother Res
Quercetin as a promising intervention for rat osteoarthritis by decreasing M1-polarized macrophages via blocking the TRPV1-mediated P2X7/NLRP3 signaling pathway. [Abstract]2024 Apr;38(4):1990-2006. PMID: 38372204 -
Neurosci Bull
Piezo1 Mediates Ultrasound-Stimulated Dopaminergic Neuron Protection via Synaptic Vesicle Recycling and Ferroptosis Inhibition. [Abstract]2025 May 29. PMID: 40439849 -
Chin Med
Novel intranasal delivery of sihosogansan demonstrates rapid antidepressant activity via GABAergic and BDNF/TrkB pathways: identification of potential bioactive quality markers. [Abstract]2025 Oct 6;20(1):162. PMID: 41047379 -
Ocul Surf
Hyperosmotic stress-induced NLRP3 inflammasome activation via the mechanosensitive PIEZO1 channel in dry eye corneal epithelium. [Abstract]2025 Jan 18:36:106-118. PMID: 39832672 -
Biochem Pharmacol
Puerarin inhibits cisplatin-induced ototoxicity in mice through regulation of TRPV1-dependent calcium overload. [Abstract]2024 Feb:220:115962. PMID: 38043717 -
NanoImpact
CdTe quantum dots induce apoptosis in RSC96 cells by disrupting calcium homeostasis and triggering subcellular structural dysfunction. [Abstract]2025 Jun 15:39:100572. PMID: 40527434 -
J Ethnopharmacol
Multitarget actions of TongXie-Ning granules against the gut dysfunctions of diarrhea-predominant irritable bowel syndrome. [Abstract]2026 Feb 28:357:120906. PMID: 41248724 -
Inflammation
Simvastatin Alleviates ConA-Induced Autoimmune Hepatitis by Inhibiting CD4+ T Cell Activation via Calcium-NFATC3 Pathway. [Abstract]2026 Jan 30;49(1):72. PMID: 41618046 -
aBIOTECH
DNA methylation-dependent epigenetic regulation of Verticillium dahliae virulence in plants. [Abstract]2023 Sep 20;4(3):185-201. PMID: 37970467 -
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Front Pharmacol
Glial cell line-derived neurotrophic factor inhibits mast-cell-like RBL-2H3 cells activation via Ca2+-mediated degranulation and Ca2+/CaMKⅡ/JNK pathway. [Abstract]2025 Nov 18:16:1697815. PMID: 41341025 -
Front Pharmacol
Resibufogenin Targets the ATP1A1 Signaling Cascade to Induce G2/M Phase Arrest and Inhibit Invasion in Glioma. [Abstract]2022 May 17;13:855626. PMID: 35656311
Fluo-4 AM purchased from MedChemExpress. Usage Cited in: Front Pharmacol. 2022 May 17;13:855626. [Abstract]
Intracellular Ca2+ is measured by Fluo-4/AM (for 30 min) with flow cytometry.
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Int Immunopharmacol
Type I and II pili mediate Streptococcus pneumoniae invasion into human blood-brain barrier-derived cells through extracellular matrix protein receptors and different endocytosis pathways. [Abstract]2025 May 30:160:114918. PMID: 40449266 -
Eur J Pharmacol
Sigma-1 receptor agonist properties that mediate the fast-onset antidepressant effect of hypidone hydrochloride (YL-0919). [Abstract]2023 May 5:946:175647. PMID: 36898424 -
Toxicology
Effects of CdTe quantum dot exposure on the calcium signaling pathway in rat dorsal root ganglion cells ND7/23. [Abstract]2025 Jun 4:154212. PMID: 40480474 -
ACS Omega
2025 Aug 21;10(34):39192-39202. PMID: 40918390 -
Microorganisms
Lsm1 Coordinates Mitochondrial Homeostasis, TORC1 Signaling, and Virulence in Candida albicans. [Abstract]2026 Mar 28;14(4):771. PMID: 42075169 -
iScience
TRPV6 is a potential regulator of bone resorption in bone loss induced by estrogen deficiency. [Abstract]2021 Oct 12;24(11):103261. PMID: 34778726 -
Cell Calcium
A NOX2-independent mechanism of Hv1 channel activation promotes inflammatory cytokine release from BV-2 microglia via intracellular Ca2+ mobilisation. [Abstract]2026 Mar:134:103122. PMID: 41581267 -
J Dermatol Sci
Thiotaurine inhibits melanoma progression by enhancing Ca2+ overload-induced cellular apoptosis. [Abstract]2025 Apr;118(1):29-37. PMID: 40189970 -
Exp Cell Res
Ano5 deficiency disturbed bone formation by inducing osteoclast apoptosis in Gnathodiaphyseal dysplasia. [Abstract]2025 Apr 1;447(1):114493. PMID: 40049314 -
Int J Cardiol
Dexmedetomidine alleviates myocardial ischemia/reperfusion-induced injury and Ca2+ overload via the microRNA-346-3p/CaMKIId axis. [Abstract]2021 Sep 1:338:185-195. PMID: 33731281 -
Mycotoxin Res
Aflatoxin B1 interferes with lncRNA-mRNA network to inhibit proliferation of bovine endometrial epithelial cells. [Abstract]2025 Dec 27;42(1):15. PMID: 41455056 -
J Dent Sci
SEC61 translocon subunit gamma enhances low-dose cisplatin-induced cancer-stem cell properties of head and neck squamous cell carcinoma via enhancing Ca2+-mediated autophagy. [Abstract]2023 Oct;18(4):1685-1692. PMID: 37799908 -
Mol Biol Rep
Aflatoxin B1 affects porcine alveolar macrophage growth through the calcium signaling pathway mediated by the ceRNA regulatory network. [Abstract]2023 Oct;50(10):8237-8247. PMID: 37572211 -
J Dig Dis
DREADDs-Based Chemogenetics Induced Slow Transit Constipation via Inhibition of Enteric Neurons. [Abstract]2025 Apr 14. PMID: 40223443 -
Biophys Chem
Fifty-hertz magnetic fields induce DNA damage through activating mPTP associated mitochondrial permeability transition in senescent human fetal lung fibroblasts. [Abstract]2024 Dec 3:318:107367. PMID: 39642656 -
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Evid Based Complement Alternat Med
2021 Jul 16;2021:6622848. PMID: 34335832 -
Oxid Med Cell Longev
Mechanosensitive Ion Channel Piezo1 Activated by Matrix Stiffness Regulates Oxidative Stress-Induced Senescence and Apoptosis in Human Intervertebral Disc Degeneration. [Abstract]2021 Feb 10;2021:8884922. PMID: 33628392
Fluo-4 AM purchased from MedChemExpress. Usage Cited in: Oxid Med Cell Longev. 2021 Feb 10;2021:8884922. [Abstract]
NP cells are incubated with 5 μM Fura-4-AM for 30 min at 37°C in the dark. Intracellular Ca2+ levels measured using the specific Ca2+-sensitive fluorescent indicator Fura-4-AM and analyzed via fluorescence microscopy.
Solvent & Solubility
DMSO : 10 mg/mL (9.12 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. The compound is unstable in solutions, freshly prepared is recommended.
Please refer to the solubility information to select the appropriate solvent. The compound is unstable in solutions, freshly prepared is recommended.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Protocol
For measuring fluorescence from cells in suspension, dilutions to 2 to 3×106 cells are made, from cultures of rat basophilic leukemia (RBL) cells. Cells are incubated in suspension in 1 μM dye (including Fluo-4 AM) for 30 min at 37°C. Cell suspensions are then transferred to cuvets for measurements of fluorescence emission intensity by spectrofluorometer[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Purity & Documentation
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Data Sheet (278 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
References
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. The compound is unstable in solutions, freshly prepared is recommended.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 0.9116 mL | 4.5581 mL | 9.1163 mL | 22.7907 mL |
| 5 mM | 0.1823 mL | 0.9116 mL | 1.8233 mL | 4.5581 mL |
Fluo-4 AM Related Classifications
HY-101896 Related Classifications
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