Fluorescein-6-isothiocyanate
(Synonyms: 6-异硫氰酸荧光素) 目录号 : GC64503Fluorescein-6-isothiocyanate 是一种荧光异构半抗原探针,对 N-Acetyl-L-Lysine、正常小鼠 IgG 和 4-4-20 的 Kd分别为 8.74、2.72 和 1.88。
Cas No.:18861-78-4
Sample solution is provided at 25 µL, 10mM.
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Fluorescein-6-isothiocyanate is a fluorescent isomeric haptenic probes with Kds of 8.74, 2.72 and 1.88 for N-Acetyl-L-Lysine, normal mouse IgG and 4-4-20, respectively[1].
[1]. Swindlehurst CA, et al. Fluorescence measurements of immune complexes of Mab 4-4-20 with isomeric haptens. Biophys J. 1991 Mar;59(3):619-28.
Cas No. | 18861-78-4 | SDF | Download SDF |
别名 | 6-异硫氰酸荧光素 | ||
分子式 | C21H11NO5S | 分子量 | 389.38 |
溶解度 | DMSO : 250 mg/mL (642.05 mM; Need ultrasonic) | 储存条件 | 4°C, 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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1 mg | 5 mg | 10 mg | |
1 mM | 2.5682 mL | 12.8409 mL | 25.6819 mL |
5 mM | 0.5136 mL | 2.5682 mL | 5.1364 mL |
10 mM | 0.2568 mL | 1.2841 mL | 2.5682 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 | ||||||||||
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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。
Analysis of 3-(2-(ethylamino)propyl)-1,2,3,4-tetrahydro-5H(1)benzopyrano (3,4-c)pyridin-5-one in plasma by liquid chromatographic column switching after derivatizing the secondary amine with Fluorescein-6-isothiocyanate
J Pharm Sci 1985 Oct;74(10):1091-4.PMID:3935773DOI:10.1002/jps.2600741015.
A high-performance liquid chromatographic (HPLC) assay has been developed to quantify the bronchodilator 3-(2-ethylamino)propyl)-1,2,3,4-tetrahydro-5H(1)benzopyrano(3,4-c)pyridi n-5-one (CI-923) (1) in human plasma. The drug and internal standard (2) were extracted from plasma (pH 10.5) with ether, and the secondary amines were derivatized with Fluorescein-6-isothiocyanate (F6ITC) (3). The derivatives were isolated from the mixture by on-line HPLC column switching and were quantified with fluorometric detection (Ex: 484 nm; Em: 518 nm). Linearity was observed over the range 0.5-50.0 ng/mL of 1, and the recovery of drug from plasma was greater than 86%. The precision of the method was 3.7-5.2% RSD based on the analysis of seeded control pools containing 0.8, 20.0 and 40.0 ng/mL of 1. The accuracy of the method was +/- 3.3% relative error for the same three pools.
Fluorescence measurements of immune complexes of Mab 4-4-20 with isomeric haptens
Biophys J 1991 Mar;59(3):619-28.PMID:1904783DOI:10.1016/S0006-3495(91)82277-9.
Relative differences in the active site environment of a monoclonal antibody when covalently bound to two isomeric haptens were studied using fluorescence quenching and lifetime measurements. Murine monoclonal antibody 4-4-20, a well-characterized high affinity antifluorescein antibody, served as the model IgG protein. Isomeric haptenic probes comparatively studied were fluorescein-5-isothiocyanate (FITC I, the immunogen) and Fluorescein-6-isothiocyanate (FITC II). In kinetic binding studies, the association rate for the interaction of 4-4-20 with FITC I was greater than 2,000 times faster than the reaction with FITC II. Fluorescence lifetimes for FITC I covalently bound to 4-4-20 were 3.89 ns and 0.37 ns, indicative of hapten bound outside and inside the active site, respectively. Fluorescence lifetime for FITC II within the active site was indistinguishable from bound FITC I, indicating that interactions with active site residues which resulted in a decreased lifetime were similar for both isomers. A decreased lifetime for active site bound FITC I was consistent with the 90-95% quenching of fluorescein fluorescence. Dynamic fluorescence quenching experiments with iodide and FITC I in the active site showed no solvent accessibility, whereas bound FITC II showed significant accessibility. These results suggest that the difference in bond angle which accompanies binding of isomer II relative to isomer I within the active site probably leads to steric constraints resulting in a more open configuration of the 4-4-20 active site.