Calcein (Fluorexon)
(Synonyms: 钙黄绿素) 目录号 : GC30082
Calcein (Fluorexon) is a fluorescent fluid phase marker used to track and visualize cellular processes such as synaptic vesicle fusion. Calcein is also the fluorophore for live cells in the commonly used Live/Dead viability assay.
Cas No.:1461-15-0,154071-48-4
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
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Cell experiment: |
Coelomocytes suspensions (ISO and CF) are separately incubated with calcein-AM (excitation and emission wavelength: 496 nm and 516 nm, respectively) at a final concentration of 200 nM, during 30 min, at 26°C. Calcein-AM is a nonfluorescent ABC transporter substrate whose intracellular accumulation is inversely proportional to ABC transporter activity. Intracellular esterases convert calcein-AM into the fluorescent dye calcein, which is not an ABC transporter substrate, thereby accumulating the dye inside the cell. Therefore, a high fluorescence signal indicates low ABC transporter activity whereas a low fluorescence signal indicates high activity. The fluorescence of samples is measured by flow cytometer. The experiment is repeated six times in duplicates. |
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References: [1]. Marques-Santos LF, et al. ABCB1 and ABCC1-like transporters in immune system cells from sea urchins Echinometra lucunter and Echinus esculentus and oysters Crassostrea gasar and Crassostrea gigas. Fish Shellfish Immunol. 2017 Sep 5;70:195-203. |
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Calcein (Fluorexon) is a fluorescent fluid phase marker used to track and visualize cellular processes such as synaptic vesicle fusion. Calcein is also the fluorophore for live cells in the commonly used Live/Dead viability assay.
[1] Brooke A Miller, et al. ACS Chem Neurosci. 2017 Oct 18;8(10):2309-2314.
| Cas No. | 1461-15-0,154071-48-4 | SDF | |
| 别名 | 钙黄绿素 | ||
| Canonical SMILES | O=C1OC2(C3=C(OC4=C2C=C(CN(CC(O)=O)CC(O)=O)C(O)=C4)C=C(O)C(CN(CC(O)=O)CC(O)=O)=C3)C5=C1C=CC=C5 | ||
| 分子式 | C30H26N2O13 | 分子量 | 622.53 |
| 溶解度 | DMSO : ≥ 100 mg/mL (160.63 mM) | 储存条件 | Store at RT, protect from light |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.6063 mL | 8.0317 mL | 16.0635 mL |
| 5 mM | 0.3213 mL | 1.6063 mL | 3.2127 mL |
| 10 mM | 0.1606 mL | 0.8032 mL | 1.6063 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Tracking the Endosomal Escape: A Closer Look at Calcein and Related Reporters
Macromol Biosci 2022 Oct;22(10):e2200167.35933579 10.1002/mabi.202200167
Crossing the cellular membrane and delivering active pharmaceuticals or biologicals into the cytosol of cells is an essential step in the development of nanomedicines. One of the most important intracellular processes regarding the cellular uptake of biologicals is the endolysosomal pathway. Sophisticated nanocarriers are developed to overcome a major hurdle, the endosomal entrapment, and delivering their cargo to the required site of action. In parallel, in vitro assays are established analyzing the performance of these nanocarriers. Among them, the release of the membrane-impermeable dye Calcein has become a popular and straightforward method. It is accessible for most researchers worldwide, allows for rapid conclusions about the release potential, and enables the study of release mechanisms. This review is intended to provide an overview and guidance for scientists applying the Calcein release assay. It comprises a survey of several applications in the study of endosomal escape, considerations of potential pitfalls, challenges, and limitations of the assay, and a brief summary of complementary methods. Based on this review, it is hoped to encourage further research groups to take advantage of the Calcein release assay for their own purposes and help to create a database for more efficient cross-correlations between nanocarriers.
Validity of Fluorexon disodium versus sodium fluorescein for use in Goldmann tonometry
Cornea 2006 Jul;25(6):679-86.17077660 10.1097/01.ico.0000214233.74603.ce
Purpose: To evaluate the safety, validity, and comfort of 0.35% Fluorexon disodium and 0.4% benoxinate (Flura-Safe) compared with the gold standard of 0.25% sodium fluorescein and 0.4% benoxinate for Goldmann applanation tonometry (GAT). Methods: This was a double-masked, randomized, crossover clinical trial. Subjects received either the standard or study formulation for GAT on visit 1 and the other formulation 1 week later. At each visit, tonometer mire quality, adequacy of fluorescence, ease of intraocular pressure (IOP) measurements, the IOP value, and anesthetizing efficacy of the formulation were assessed. Subjects graded general comfort, soreness and irritation, and burning and stinging of each formulation at 1 and 5 minutes after drop instillation. Results: Sixty-seven subjects completed the study. The mean IOP was 13.9 +/- 2.7 with Fluorexon and 13.9 +/- 2.8 mm Hg with fluorescein OD and 14.0 +/- 2.8 with Fluorexon and 13.9 +/- 2.5 mm Hg with fluorescein OS. The measurements with the 2 formulations were highly correlated for OD and OS, and the differences between the 2 measurements were not clinically significant. There was also no significant difference between the 2 drops in mire clarity, adequacy of fluorescence, or corneal anesthesia. However, Fluorexon was statistically more comfortable (P = 0.039) and caused less stinging and burning (P = 0.014) at 1 minute versus the fluorescein formulation. Conclusion: Not only was the new Fluorexon product accurate and effective in GAT, it was also statistically more comfortable and had a less stinging and burning effect at 1 minute after drop instillation than the traditional fluorescein formulation. Because Fluorexon is less likely to stain soft contact lenses, this may be the dye-anesthetic formulation of choice for practices that routinely perform GAT.
Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products
Nat Protoc 2008;3(5):877-82.18451795 10.1038/nprot.2008.57
As the human genome is decoded and its involvement in diseases is being revealed through postgenome research, increased adoption of genetic testing is expected. Critical to such testing methods is the ease of implementation and comprehensible presentation of amplification results. Loop-mediated isothermal amplification (LAMP) is a simple, rapid, specific and cost-effective nucleic acid amplification method when compared to PCR, nucleic acid sequence-based amplification, self-sustained sequence replication and strand displacement amplification. This protocol details an improved simple visual detection system for the results of the LAMP reaction. In LAMP, a large amount of DNA is synthesized, yielding a large pyrophosphate ion by-product. Pyrophosphate ion combines with divalent metallic ion to form an insoluble salt. Adding manganous ion and Calcein, a fluorescent metal indicator, to the reaction solution allows a visualization of substantial alteration of the fluorescence during the one-step amplification reaction, which takes 30-60 min. As the signal recognition is highly sensitive, this system enables visual discrimination of results without costly specialized equipment. This detection method should be helpful in basic research on medicine and pharmacy, environmental hygiene, point-of-care testing and more.
Ultrasound-induced Calcein release from eLiposomes
Ultrasound Med Biol 2012 Dec;38(12):2163-73.23062373 10.1016/j.ultrasmedbio.2012.08.001
Ultrasound is explored as a method of inducing the release of encapsulated materials from eLiposomes, defined as liposomes containing emulsion droplets. Emulsions were formed using perfluorohexane and perfluoropentane. eLiposomes were formed by folding interdigitated lipid sheets into closed vesicles around the emulsion droplets. Cryogenic transmission electron microscopy was used to verify droplet encapsulation. Self-quenched Calcein was also encapsulated inside the vesicles. A fluorometer was used to measure baseline fluorescence, Calcein release after ultrasound exposure, and total release from the vesicles. eLiposome samples released 3 to 5 times more of the encapsulated Calcein than did controls when exposed to 20-kHz ultrasound. Calcein release increased with exposure time and intensity of ultrasound. eLiposomes with large (400 nm) droplets produced more Calcein release than small (100 nm) droplets. These observations suggest that the emulsions are vaporized by ultrasound and that the Laplace pressure in the emulsions has an effect on droplet vaporization.
Calcein Release from Cells In Vitro via Reversible and Irreversible Electroporation
J Membr Biol 2018 Feb;251(1):119-130.29143077 10.1007/s00232-017-0005-8
The aim of this study was to investigate the dependence of Calcein extraction and cell viability on the parameters of pulsed electric field (PEF). Two different approaches concerning PEF parameters were investigated: (1) extraction efficiency and cell viability dependence on pulse number, exploiting 1200 V/cm 100 µs duration high voltage (HV) electric pulses and (2) extraction efficiency and cell viability dependence on the pulses with different duration (44-400 µs) and electric field strength (600-1800 V/cm) that result in the same amount of electric field energy delivered to Chinese hamster ovary cells. Extraction efficiency was evaluated as a percentage ratio of Calcein fluorescence intensity prior and after PEF treatment. Cell viability was evaluated using PI test and cell clonogenic assay. Moreover, Calcein release dynamics from cells after 600 V/cm 400 µs, 1200 V/cm 100 µs, and 1800 V/cm 44 µs was evaluated. Our results show that HV pulses induce instant Calcein extraction due to reversible electroporation; however, subsequent Calcein leakage over time was only observed when 9 HV pulses of 1800 V/cm 44 µs were used. The increased number of pulses resulted in more efficient total Calcein extraction. With the same total energy delivered via electric pulses, the increase of Calcein extraction efficiency was more dependent on pulse strength rather than pulse duration. The highest Calcein extraction efficiency (84.5 ± 7.4%) was obtained using 9 electric field pulses of 1800 V/cm, 44 µs at 1 Hz. Furthermore, the extraction efficiency can be significantly enhanced if external mechanical stress (pipetting) is applied to cells. Cell viability was determined to be dependent on different PEF exposure parameters. It varied from 96.8 ± 4.8 to 31.2 ± 8.9%, implying the possibility to adjust PEF parameter combinations to maintain high cell viability.