Calcein AM | AAT Bioquest

Calcein, AM

CAS 148504-34-1

Calcein AM readily passes through the cell membrane of viable cells because of its enhanced hydrophobicity as compared to calcein. The acetomethoxy (AM) derivate of calcein (calcein AM) is widely used for labeling live cells as it can be transported through the cellular membrane into live cells. The AM ester groups mask the part of the molecule that chelates calcium. Upon transporting into live cells cellular esterases cut off the AM groups, the molecule binds to calcium within cell (resulting in acquiring strong green fluorescence), and gets trapped inside. As dead cells lack esterases, only live cells are marked. This feature makes it very useful for testing of cell viability and for short-term marking of cells. Compared with other live cell-labeling reagents (such as BCECF-AM and carboxy-fluorescein diacetate), calcein-AM is the most suitable fluorescent probe for staining viable cells because of its low cytotoxicity. Calcein does not significantly affect cellular functions such as proliferation or chemotaxis of lymophocyte. In addition, viability assays using calcein are reliable and correlate well with the standard 51Cr-release assay.

Simultaneous imaging of live and apoptotic HeLa cells labeled using calcein AM (Cat No. 22002) and Annexin V-iFluor® 647 conjugate (Cat No. 20074).

Protocol

SDS

Catalog	Size	Price	Quantity
22002	1 mg	Price

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Calcein, AM *CAS 148504-34-1* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	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

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	994.86 g/mol	=	mL	x	mM	=	mmol

Citations

View all 55 citations: Citation Explorer

Liver-on-Micropillar: A Humanized, Animal-Free Platform for High-Throughput Assessment of Drug-Induced Liver Injury

Authors:

Ali, Ahmed SMSM and Wu, Dongwei and R{\"o}hrs, Viola and Nguyen, Duy Bao Tran and Ziersch, Mathias and Braeuning, Albert and Kurreck, Jens

Journal:

Biofabrication (2025)

Elucidating the distinctive regulatory effects and mechanisms of active compounds in Salvia miltiorrhiza Bunge via network pharmacology: Unveiling their roles in the modulation of platelet activation and thrombus formation

Authors:

Zhang, Ying and Xin, Guang and Zhou, Qilong and Yu, Xiuxian and Feng, Lijuan and Wen, Ao and Zhang, Kun and Wen, Tingyu and Zhou, Xiaoli and Wu, Qiuling and others,

Journal:

Toxicology and Applied Pharmacology (2024): 116871

Reinforced Immunogenic Endoplasmic Reticulum Stress and Oxidative Stress via an Orchestrated Nanophotoinducer to Boost Cancer Photoimmunotherapy

Authors:

Yang, Zhenzhen and Teng, Yulu and Lin, Meng and Peng, Yiwei and Du, Yitian and Sun, Qi and Gao, Datong and Yuan, Quan and Zhou, Yu and Yang, Yiliang and others,

Journal:

ACS nano (2024)

Type I collagen and fibromodulin enhance the tenogenic phenotype of hASCs and their potential for tendon regeneration

Authors:

Tu, Tian and Shi, Yuan and Zhou, Boya and Wang, Xiaoyu and Zhang, Wenjie and Zhou, Guangdong and Mo, Xiumei and Wang, Wenbo and Wu, Jinglei and Liu, Wei

Journal:

npj Regenerative Medicine (2023): 67

Hyperglycemia-regulated tRNA-derived fragment tRF-3001a propels neurovascular dysfunction in diabetic mice

Authors:

Zhu, Jun-Ya and Yao, Wen and Ni, Xi-Sen and Yao, Mu-Di and Bai, Wen and Yang, Tian-Jing and Zhang, Zi-Ran and Li, Xiu-Miao and Jiang, Qin and Yan, Biao

Journal:

Cell Reports Medicine (2023)

References

View all 84 references: Citation Explorer

Functional evidence that the self-renewal gene NANOG regulates esophageal squamous cancer development

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)

Localized functional chemical stimulation of TE 671 cells cultured on nanoporous membrane by calcein and acetylcholine

Authors:

Zibek S, Stett A, Koltay P, Hu M, Zengerle R, Nisch W, Stelzle M.

Journal:

Biophys J. (2006)

A vaccination and challenge model using calcein marked fish

Authors:

Klesius PH, Evans JJ, Shoemaker CA, Pasnik DJ.

Journal:

Fish Shellfish Immunol (2006): 20

Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viability and aging

Authors:

Bratosin D, Mitrofan L, Palii C, Estaquier J, Montreuil J.

Journal:

Cytometry A (2005): 78

Cytotoxic effects of 100 reference compounds on Hep G2 and HeLa cells and of 60 compounds on ECC-1 and CHO cells. I mechanistic assays on ROS, glutathione depletion and calcein uptake

Authors:

Schoonen WG, Westerink WM, de Roos JA, Debiton E.

Journal:

Toxicol In Vitro (2005): 505

Physical properties

Molecular weight	994.86
Solvent	DMSO

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	81000
Excitation (nm)	501
Emission (nm)	521

Storage, safety and handling

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
CAS	148504-34-1

Instrument settings

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

Documents

Protocol
Safety Data Sheet
Certificate of Origin
Certificate of Analysis
Brochure: Cell Viability & Proliferation
Flyer: Calcein Dyes

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Page updated on September 13, 2024