Calcein-AM (Calcein acetoxymethyl ester) (Synonyms: Calcein Acetoxymethyl ester, NSC 689290) |
Catalog No.GC34061
One-Click Copy Product Info
|
Calcein-AM is a highly lipophilic vital dye that rapidly enters viable cells.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 148504-34-1
Sample solution is provided at 25 µL, 10mM.
- Embo Mol Med (2024):1-24.PMID:39148004
- Int J Nanomed: (2026): 611830.PMID:42615011
- Int J Biol Sci 17.11 (2021):2703.PMID:34345202
- Nat Product Bioprosp 14.1 (2024):1-13.PMID:38780674
- Iscience (2025).
- Phytomedicine 105 (2022):154330.PMID:35905566
- Molecules 28.8 (2023):3389.PMID:37110625
- Phytomedicine (2024):155463.
- J Cancer 15.14 (2024):4577-4590.
Calcein-AM is a highly lipophilic vital dye that rapidly enters viable cells. It is converted by intracellular esterases to calcein that produces an intense green (530-nm) signal, and is retained by cells with intact plasma membrane.[1]
In vitro experiment it shown that calcein-AM assay used to assess human RBC viability after incubation (37°C for 3 and 20 h) in the presence of Ca2+ (2.5 mM) and ionophore A 23187 (0.5 μM).[1] 0.05 μM was the optimal concentration of CAM (Calcein-AM) for staining effector cells by testing 0.05, 0.1, 0.2, and 0.4 μM. Using 0.05 μM CAM to stain the PBMCs and expanded NK cells from three normal volunteers, the results demonstrated that there is no significant decrease in cytotoxicity and CAM staining had no significant effect on human NK cell activity in PBMCs or in expanded NK cells.[2] In vitro, 50μm calcein AM's fluorescent signal of 1 x lo5 lymphocytes was close to the saturation level, while the signal emitted by lymphocytes labeled with 20μm calcein AM was only slightly lower. [3] Calcein-AM has cytotoxic activity against human tumor cell lines (such as the human lymphoma U-937-GTB) at low concentrations (2.5 ug/ml).[4] In vitro experiment it demonstrated that in the mixed macrophages and THP-1 cells (5x105 cells/ml), Calcein-AM (2 µM)/propidium Iodide (PI) (4.5 µM) staining assay Calcein-AM/PI double staining was used to quantify the number of living and dead cells as a cell death assay.[5] In addition, The cells in OA chondrocytes were seeded in 24-well plates (2 × 104 cells/well), cultured for 4 h, the cells were treated with 5 μL Calcein-AM (2 μM) and 5 μL PI (2 μM) at 37°C in conditions void of light for 30 min, and then analyzed under a fluorescence microscope.[6]
References:
[1].Bratosin D, et al. Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viability and aging. Cytometry A. 2005 Jul;66(1):78-84.
[2].Jang YY, et al. An improved flow cytometry-based natural killer cytotoxicity assay involving calcein AM staining of effector cells. Ann Clin Lab Sci. 2012 Winter;42(1):42-9.
[3].Braut-Boucher F, et al. A non-isotopic, highly sensitive, fluorimetric, cell-cell adhesion microplate assay using calcein AM-labeled lymphocytes. J Immunol Methods. 1995 Jan 13;178(1):41-51.
[4].Liminga G, et al. Cytotoxic effect of calcein acetoxymethyl ester on human tumor cell lines: drug delivery by intracellular trapping. Anticancer Drugs. 1995 Aug;6(4):578-85.
[5].Xiang N, et al. Gardnerella vaginalis induces NLRP3 inflammasome-mediated pyroptosis in macrophages and THP-1 monocytes. Exp Ther Med. 2021 Oct;22(4):1174.
[6].Zhang L, et al. MicroRNA-140-5p represses chondrocyte pyroptosis and relieves cartilage injury in osteoarthritis by inhibiting cathepsin B/Nod-like receptor protein 3. Bioengineered. 2021 Dec;12(2):9949-9964.
| Cell experiment [1]: | |
|
Cell lines |
EPC cells |
|
Preparation Method |
Uptake kinetics of EPC cells loaded with 5 µM calcein AM seeded at 1x105 cells well-1 and cultured at 15 °C. Calcein AM uptake was measured as fluorescence intensity (FI). |
|
Reaction Conditions |
5 µM at 15 °C, 1-8h |
|
Applications |
Uptake kinetics showed that for EPC cells seeded at a density of 1x105 cells well-1 calcein AM labelling increased throughout the 7 h tested. |
|
References: |
|
| Cas No. | 148504-34-1 | SDF | |
| Synonyms | Calcein Acetoxymethyl ester, NSC 689290 | ||
| Chemical Name | tetrakis(acetoxymethyl) 2,2',2'',2'''-(((3',6'-diacetoxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-4',5'-diyl)bis(methylene))bis(azanetriyl))tetraacetate | ||
| Canonical 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 | ||
| Formula | C46H46N2O23 | M.Wt | 994.86 |
| Solubility | 10 mg/mL in EtOH, MeOH, DMSO, DMF with gentle heating (37°C) for 10 minutes. | Storage | -20°C, protect from light |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.0052 mL | 5.0258 mL | 10.0517 mL |
| 5 mM | 201 μL | 1.0052 mL | 2.0103 mL |
| 10 mM | 100.5 μL | 502.6 μL | 1.0052 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
-
Related Biological Data
Construction of breast cancer organoid in vitro for drug testing via printing homo-MA and hetero-MA bioinks:d) cell live/dead staining of the core and shell part of breast cancer organoid after exposure to cisplatin (500 umol L−1) and paclitaxel (100 nmol L−1) for 24 and 48h.
Cell viability was tested by Calcein-AM/PI Double Staining Kit. 2 mL of PBS solution containing 0.1 vol% Calcein AM(GlpBio) and 0.1 vol% PI was added into the samples and incubated for 30 min at 37 °C.
Small Methods (2024): 2400223. PMID: 38602202 IF: 12.4002 -
Related Biological Data
MSC coculture inhibits neuronal ferroptosis through mitochondrial transfer.(K) Flow cytometry analysis of LIP in Ht22 cells from culture alone, coculture with MSCs or MSC coculture combined with Cyto D treatment after Rsl3 stimulation for 24 h. Cells were incubated by Calcein-AM (FITC channel) and treated with or without Deferiprone.
After trypsinization, cells (6-well dishes) were washed twice with PBS and then incubated with 1 μM Calcein-AM(GlpBio) at 37°C for 15 min.
Redox Biology, 2023: 102871. PMID: 37699320 IF: 11.3997 -
Related Biological Data
PEF inhibits ferroptosis in C. elegans models of AD.(L) Representative images of CL4176 worms stained with Calcein-AM reagent in the presence or absence of PEF (800 μg/mL) and DFO (100 μM). (M) The bar chart shows the relative Calcein-AM intensity in CL4176 worms.
Briefly, after treatment, worms were collected and co-cultured in M9 with 0.05 μg/mL Calcein-AM(GlpBio) for 1 hour, followed by three washes in 1 mL M9.
Phytomedicine (2024): 155463. PMID: 38452694 IF: 7.8996 -
Related Biological Data
NAR reduced cellular iron concentration. (A) Chondrocytes were treated with IL-1β and FAC, with or without various concentrations of NAR for 24 hr, and then stained with a Calcein-AM fluorescent probe for 15min.
Chondrocytes were treated with IL-1β and FAC, with or without various concentrations of NAR for 24 hr, and then stained with a Calcein-AM(GlpBio) fluorescent probe for 15min.
Phytomedicine (2022): 154330. PMID: 35905566 IF: 6.6563 -
Related Biological Data
Tagitinin C induces ferroptosis in HCT116 cells.(G) Quantification of cellular LIP levels using the calcein-AM (C-AM) method. The mean fluorescence intensity (MFI) of C-AM is subtracted from the MFI of C-AM treated with DFO.
After trypsinization, the cells were washed twice with PBS followed by incubation of 2 µM calcein-acetoxymethyI ester (GLPBIO) for 30 min at 37 °C.
Int J Biol Sci 17.11 (2021): 2703-2717. PMID: 34345202 IF: 6.58
Average Rating: 5 (Based on Reviews and 1 reference(s) in Google Scholar.)