Calcein Blue
(Synonyms: 钙黄绿素蓝) 目录号 : GC43117
A membrane-impermeant fluorescent dye
Cas No.:54375-47-2
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Cell experiment: | The recommended final working concentrations is usually between 1 and 10 μM to minimize potential artifacts. Generally, 15 minutes to 1 hour is sufficient for cellular uptake and processing of the dyes. A stock concentration of 5 mM 4-methylumbelliferone-8- methyliminodiacetic acid, commonly known as calcein blue is prepared in 0.1 M potassium hydroxide, and the pH is neutralized using 0.1 N hydrochloric acid. The calcein blue stock prepared is diluted to 200 μM (working stock) in DPBS and stored for further use. A concentration of 10 μM calcein blue is prepared from the working stock by diluting in DPBS. From this, 200 μL is added to a 96-well plate and the excitation/emission (Ex./Em.) maximum is determined using the micro-plate reader[1]. |
References: [1]. Sankaranarayanan R, et al. A new fluorimetric method for the detection and quantification of siderophores using Calcein Blue, with potential as a bacterial detection tool. Appl Microbiol Biotechnol. 2015 Mar;99(5):2339-49. | |
Calcein Blue is a short-term, blue-fluorescent dye.
Calcein blue AM is only weakly fluorescent (excitation/emission maxima 322/435 nm). Upon cleavage of the AM esters by intracellular esterases, this tracer becomes relatively polar and is retained by cells for several hours. In addition, its fluorescence intensity increases and its fluorescence spectra shift to longer wavelengths, with excitation/emission maxima of 360/449 nm. The fluorescence of Calcein Blue is known to be quenched in the presence of iron(III); if a stronger chelator removes this ion from the fluorophore, the fluorescence of the fluorophore is regained[1]. A novel fluorimetric assay for dopamine using calcein blue (CB) complexed with Fe2 ion as a chemical sensor is described. The fluorescence arising from calcein blue of the calcein blue–Fe2 complex is quenched by the Fe2 ion. When dopamine is added to a solution of the calcein blue–Fe2 complex, a dopamine–Fe2 complex is formed as the result of a ligand exchange reaction between calcein blue and dopamine which permits the fluorescence from calcein blue to be recovered. The fluorescence intensity at the wavelength of 440 nm (at the excitation wavelength of 340 nm) is found to be proportional to the concentration of the dopamine added to the calcein blue–Fe2 complex solution, which permits dopamine to be quantitatively determined[2].
References:
[1]. Sankaranarayanan R, et al. A new fluorimetric method for the detection and quantification of siderophores using Calcein Blue, with potential as a bacterial detection tool. Appl Microbiol Biotechnol. 2015 Mar;99(5):2339-49.
[2]. Seto D, et al. A simple and selective fluorometric assay for dopamine using a calcein blue-Fe2+ complex fluorophore. Talanta. 2012 May 30;94:36-43.
| Cas No. | 54375-47-2 | SDF | |
| 别名 | 钙黄绿素蓝 | ||
| Canonical SMILES | OC1=CC=C(C(C)=CC(O2)=O)C2=C1CN(CC(O)=O)CC(O)=O | ||
| 分子式 | C15H15NO7 | 分子量 | 321.3 |
| 溶解度 | DMSO : 1 mg/mL (3.11 mM; Need ultrasonic); H2O : < 0.1 mg/mL (insoluble) | 储存条件 | Store at -20°C |
| 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 | 3.1124 mL | 15.5618 mL | 31.1236 mL |
| 5 mM | 0.6225 mL | 3.1124 mL | 6.2247 mL |
| 10 mM | 0.3112 mL | 1.5562 mL | 3.1124 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。
On the properties of Calcein Blue
Talanta 1974 Dec;21(12):1221-9.PMID:18961591DOI:10.1016/0039-9140(74)80143-8.
A satisfactory method for the preparation of Calcein Blue has been devised. Elemental analysis, equivalent weight by neutralization, and the NMR spectrum show the compound to be 4-methylumbelliferone-8-methyleneiminodiacetic acid.0.25H(2)O. The ultraviolet absorbance and fluorescence have been studied as a function of pH and, combined with potentiometric titration and solubility date, have yielded for the acid dissociation constants the values pK(1) = 3.0, pK(2) = 6.9, and pK(3) = 11.3. These acid functions are identified respectively as carboxyl, phenol, and ammonium ion, the free Calcein Blue being a zwitter-ion. Calcein Blue fluoresces in both acidic and basic solution when excited at a suitable wavelength. The fluorescence of the doubly-charged anion formed on the neutralization of the phenol group, when excited at 360 nm, reaches a maximum at pH 9, and decreases to zero with the neutralization of the ammonium ion; the wavelength of maximum emission is 455 nm. In the presence of calcium, the fluorescence increases with alkalinity up to pH 9 and then remains constant. The calcium derivative is a 1:1 compound, formation constant 10(7.1). The fluorescence of Calcein Blue at all pH values is quenched by copper(II). The calcium derivative is changed on standing in highly alkaline solution, presumably by ring opening, to another fluorescent material; thus Calcein Blue, although satisfactory as an indicator, is not useful for the direct fluorometric determination of calcium.
Methyl Calcein Blue and other analogues of Calcein Blue
Talanta 1974 Nov;21(11):1193-202.PMID:18961582DOI:10.1016/0039-9140(74)80102-5.
4-Methylumbelliferone-8-methylenesarcosine (Methyl Calcein Blue) and four related metallofluorochromic indicators derived from umbelliferone, 4-methylumbelliferone, and 4-methylesculetin by condensation with formaldehyde and iminodiacetic acid or glycine have been synthesized, the structures established, the absorbance and fluorescence measured as functions of pH, and the reactions with copper(II) and calcium studied with attention to the effects on fluorescence. All of the compounds display a maximum fluorescence at ph about 9. The fluorescence of each is quenched by copper(II). The calcium derivatives of the compounds derived from the umbelliferones and iminodiacetic acid fluoresce at high ph but those from the umbelliferones and glycine or sarcosine do not. At high pH, 4-methylesculetin and the amino-acids derived from it do not fluoresce either alone or in the prescence of calcium.
Detecting microdamage in bone
J Anat 2003 Aug;203(2):161-72.PMID:12924817DOI:10.1046/j.1469-7580.2003.00211.x.
Fatigue-induced microdamage in bone contributes to stress and fragility fractures and acts as a stimulus for bone remodelling. Detecting such microdamage is difficult as pre-existing microdamage sustained in vivo must be differentiated from artefactual damage incurred during specimen preparation. This was addressed by bulk staining specimens in alcohol-soluble basic fuchsin dye, but cutting and grinding them in an aqueous medium. Nonetheless, some artefactual cracks are partially stained and careful observation under transmitted light, or epifluorescence microscopy, is required. Fuchsin lodges in cracks, but is not site-specific. Cracks are discontinuities in the calcium-rich bone matrix and chelating agents, which bind calcium, can selectively label them. Oxytetracycline, alizarin complexone, calcein, Calcein Blue and xylenol orange all selectively bind microcracks and, as they fluoresce at different wavelengths and colours, can be used in sequence to label microcrack growth. New agents that only fluoresce when involved in a chelate are currently being developed--fluorescent photoinduced electron transfer (PET) sensors. Such agents enable microdamage to be quantified and crack growth to be measured and are useful histological tools in providing data for modelling the material behaviour of bone. However, a non-invasive method is needed to measure microdamage in patients. Micro-CT is being studied and initial work with iodine dyes linked to a chelating group has shown some promise. In the long term, it is hoped that repeated measurements can be made at critical sites and microdamage accumulation monitored. Quantification of microdamage, together with bone mass measurements, will help in predicting and preventing bone fracture failure in patients with osteoporosis.
A new fluorimetric method for the detection and quantification of siderophores using Calcein Blue, with potential as a bacterial detection tool
Appl Microbiol Biotechnol 2015 Mar;99(5):2339-49.PMID:25634020DOI:10.1007/s00253-015-6411-x.
The presence of microorganisms in biological fluids like urine and blood is an indication of vulnerability to infections. Iron is one of the important micronutrients required for bacterial growth. In an iron-deficit environment, bacteria release high-affinity iron-chelating compounds called siderophores which can be used as non-invasive target molecules for the detection of such pathogens. However, only limited reagents and procedures are available to detect the presence of these organic molecules. The present study aims at detecting the presence of siderophores in the iron-depleted media, exploiting the reversible quenching of Calcein Blue and iron(III) complex. The fluorescence of Calcein Blue is known to be quenched in the presence of iron(III); if a stronger chelator removes this ion from the fluorophore, the fluorescence of the fluorophore is regained. This behaviour of the fluorophore was exploited to detect and quantify siderophores down to 50 and 800 nM equivalent of standard siderophore, deferroxamine mesylate (desferal) in Dulbecco's PBS and siderophore quantification (SPQ) medium, respectively. The siderophores released by pathogens, equivalent to standard desferal, were in the range of 1.29 to 5.00 μM and those for non-pathogens were below 1.19 μM. The simple, sensitive and cost-effective method performed in a 96-well plate was able to detect and quantify iron chelators within 7-8 h of incubation.
A simple and selective fluorometric assay for dopamine using a calcein blue-Fe2+ complex fluorophore
Talanta 2012 May 30;94:36-43.PMID:22608411DOI:10.1016/j.talanta.2012.02.025.
A novel fluorimetric assay for dopamine using Calcein Blue (CB) complexed with Fe(2+) ion as a chemical sensor is described. The fluorescence arising from CB of the CB-Fe(2+) complex is quenched by the Fe(2+) ion. When dopamine is added to a solution of the CB-Fe(2+) complex, a dopamine-Fe(2+) complex is formed as the result of a ligand exchange reaction between CB and dopamine which permits the fluorescence from CB to be recovered. The fluorescence intensity at the wavelength of 440 nm (at the excitation wavelength of 340 nm) was found to be proportional to the concentration of the dopamine added to the CB-Fe(2+) complex solution, which permits dopamine to be quantitatively determined. The selectivity for dopamine in the presence of other catecholamines and related compounds was good. The calibration curve for dopamine, determined using experimental data was successfully simulated based on the equilibrium of the ligand exchange reaction between CB and dopamine. The working range is from 50 μM to 1mM and the limit of detection and limit of quantization are ca 10 μM and 50 μM, respectively. The assay is simple and economical, compared with conventional methods such as an enzyme-linked immunosorbent assay (ELISA).