Calcein, AM *UltraPure grade* *CAS 148504-34-1*
Price | |
Catalog Number | |
Unit Size | |
Quantity |
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Molecular weight | 994.86 |
Solvent | DMSO |
Extinction coefficient (cm -1 M -1) | 81000 |
Excitation (nm) | 501 |
Emission (nm) | 521 |
Certificate of Origin | Download PDF |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12352200 |
Calcein, AM *CAS 148504-34-1* |
Overview | ![]() ![]() |
CAS 148504-34-1 | Molecular weight 994.86 | Extinction coefficient (cm -1 M -1) 81000 | Excitation (nm) 501 | Emission (nm) 521 |
Platform
Flow cytometer
Excitation | 488 nm laser |
Emission | 530/30 nm filter |
Instrument specification(s) | FITC channel |
Fluorescence microscope
Excitation | FITC filter set |
Emission | FITC filter set |
Recommended plate | Black wall/clear bottom |
Fluorescence microplate reader
Excitation | 490 |
Emission | 525 |
Cutoff | 515 |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Bottom read mode |
Example protocol
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Prepare a 2 to 5 mM stock solution of Calcein AM in high-quality, anhydrous DMSO.
Note: When reconstituted in DMSO, Calcein AM is a clear, colorless solution.
PREPARATION OF WORKING SOLUTION
Prepare a Calcein AM working solution of 1 to 10 µM in the buffer of your choice (e.g., Hanks and Hepes buffer). For most cell lines, Calcein AM at the final concentration of 4 to 5 µM is recommended. The exact concentration of indicators required for cell loading must be determined empirically.
Note: The nonionic detergent Pluronic® F-127 can be used to increase the aqueous solubility of AM esters. In the staining buffer, the final Pluronic® F-127 concentration should be approximately 0.02%. A variety of Pluronic® F-127 products can be purchased from AAT Bioquest. Avoid long-term storage of AM esters in the presence of Pluronic® F-127.
Note: If your cells contain organic anion-transporters, probenecid (1–2.5 mM) or sulfinpyrazone (0.1–0.25 mM) may be added to the working solution to reduce leakage of the de-esterified indicators. A variety of ReadiUse™ Probenecid products, Including water-soluble, sodium salt, and stabilized solutions, can be purchased from AAT Bioquest.
SAMPLE EXPERIMENTAL PROTOCOL
- Prepare cells for imaging.
Remove the cell culture medium and wash cells once with serum-free buffer to remove any remaining media.
Note: Serum in cell culture media may contain esterase activity, which can increase background interference.
Add Calcein AM working solution to the culture.
- Incubate cells at 37 °C for 30 to 60 minutes.
- Replace the dye working solution with HHBS or buffer of your choice (containing an anion transporter inhibitor, such as 1 mM probenecid, if applicable) to remove any excess probes.
- Measure the fluorescence intensity using either a fluorescence microscope equipped with a FITC filter set, a flow cytometer equipped with a blue laser and a 530/30 nm filter (FITC channel), or a fluorescence plate reader at Ex/Em = 490/525 nm cutoff 515 nm.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 100.517 µL | 502.583 µL | 1.005 mL | 5.026 mL | 10.052 mL |
5 mM | 20.103 µL | 100.517 µL | 201.033 µL | 1.005 mL | 2.01 mL |
10 mM | 10.052 µL | 50.258 µL | 100.517 µL | 502.583 µL | 1.005 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Spectrum

Spectral properties
Extinction coefficient (cm -1 M -1) | 81000 |
Excitation (nm) | 501 |
Emission (nm) | 521 |
Images
Citations
Authors: Wolff, Anne and Frank, Marcus and Staehlke, Susanne and Springer, Armin and Hahn, Olga and Meyer, Juliane and Peters, Kirsten
Journal: Biomedicines (2023): 1049
Authors: Cheremkhina, Maria and Klein, Sarah and Babendreyer, Aaron and Ludwig, Andreas and Schmitz-Rode, Thomas and Jockenhoevel, Stefan and Cornelissen, Christian G and Thiebes, Anja Lena
Journal: Micromachines (2023): 575
Authors: Geraets, EKW
Journal: (2023)
Authors: Thiebes, Anja Lena and Klein, Sarah and Zingsheim, Jonas and M{\"o}ller, Georg H and G{\"u}rzing, Stefanie and Reddemann, Manuel A and Behbahani, Mehdi and Jockenhoevel, Stefan and Cornelissen, Christian G
Journal: Pharmaceutics (2022): 2421
Authors: Bielfeldt, Meike and Rebl, Henrike and Peters, Kirsten and Sridharan, Kaarthik and Staehlke, Susanne and Nebe, J Barbara
Journal: Biomedical Materials \& Devices (2022): 1--16
Authors: Amsri, Artid and Srichairatanakool, Somdet and Teerawutgulrag, Aphiwat and Youngchim, Sirida and Pongpom, Monsicha
Journal: Journal of Fungi (2022): 1183
Authors: Samie, Muhammad and Khan, Ather Farooq and Rahman, Saeed Ur and Iqbal, Haffsah and Yameen, Muhammad Arfat and Chaudhry, Aqif Anwar and Galeb, Hanaa A and Halcovitch, Nathan R and Hardy, John G
Journal: International Journal of Biological Macromolecules (2022)
Authors: Schauer, Tim and Djurhuus, Sissal Sigmundsd{\'o}ttir and Simonsen, Casper and Brasso, Klaus and Christensen, Jesper Frank
Journal: Brain, behavior, \& immunity-health (2022): 100508
Authors: Munguia-Lopez, Jose G and Jiang, Tao and Ferlatte, Audrey and Fajardo-Diaz, Juan L and Munoz-Sandoval, Emilio and Tran, Simon D and Kinsella, Joseph M
Journal: Advanced NanoBiomed Research (2021): 2100104
Authors: G{\"u}rzing, Stefanie and Klein, Sarah and M{\"o}ller, Georg and Thiebes, Anja Lena and Cornelissen, Christian Gabriel and Reddemann, Manuel Armin
Journal: Atomization and Sprays (2021)
References
Authors: Li, Deng and Xiang, Xiaocong and Yang, Fei and Xiao, Dongqin and Liu, Kang and Chen, Zhu and Zhang, Ruolan and Feng, Gang
Journal: Biochemical and Biophysical Research Communications (2017)
Authors: Zibek S, Stett A, Koltay P, Hu M, Zengerle R, Nisch W, Stelzle M.
Journal: Biophys J. (2006)
Authors: Klesius PH, Evans JJ, Shoemaker CA, Pasnik DJ.
Journal: Fish Shellfish Immunol (2006): 20
Authors: Bratosin D, Mitrofan L, Palii C, Estaquier J, Montreuil J.
Journal: Cytometry A (2005): 78
Authors: Schoonen WG, Westerink WM, de Roos JA, Debiton E.
Journal: Toxicol In Vitro (2005): 505
Authors: Iwanowicz LR, Densmore CL, Ottinger CA.
Journal: Fish Shellfish Immunol (2004): 127
Authors: Uggeri J, Gatti R, Belletti S, Sc and roglio R, Corradini R, Rotoli BM, Orl and ini G., undefined
Journal: Histochem Cell Biol (2004): 499
Authors: Mueller H, Kassack MU, Wiese M.
Journal: J Biomol Screen (2004): 506
Authors: Goto T, Kajiwara H, Yoshinari M, Fukuhara E, Kobayashi S, Tanaka T.
Journal: Biomaterials (2003): 3885
Authors: Tokudome Y, Sugibayashi K.
Journal: Biol Pharm Bull (2003): 1508