5-IAF (5-Iodoacetamidofluorescein)
(Synonyms: 5-碘乙酰氨基荧光素; 5-Iodoacetamidofluorescein) 目录号 : GC34180
A fluorescent probe for labeling proteins
Cas No.:63368-54-7
Sample solution is provided at 25 µL, 10mM.
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5-IAF is a fluorescent probe for labeling proteins.1 It has been used to label proteins through a reaction with sulfhydryl groups. 5-IAF displays excitation/emission maxima of 491/518 nm, respectively, which can shift during protein labeling or changes in hydrophobicity and can be quenched by potassium iodide. 5-IAF has been used to monitor ligand binding and conformational changes of the Na+/K+-ATPase.2
1.Hartig, P.R., Bertrand, N.J., and Sauer, K.5-Iodoacetamidofluorescein-labeled chloroplast coupling factor 1: Conformational dynamics and labeling-site characterizationBiochemistry16(19)4275-4282(1977) 2.Kapakos, J.G., and Steinberg, M.Ligand binding to (Na,K)-ATPase labeled with 5-iodoacetamidofluoresceinJ. Biol. Chem.261(5)2084-2089(1986)
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Kinase experiment: |
HepG2 cells are lysed in 1 mL 2% acetonitrile in water, and transfered to a 1.5 mL centrifuge tube. An additional 200 µL 2% acetonitrile is used to rinse the dish and combined with the cell extract. To measure the free GSH concentration, three aliquots of 80 µL cell extract are immediately transferred to 0.5 mL centrifuge tubes after a quick mix. Then 20 µL 50 µM NAC is added, followed by the addition of 100 µL 400 µM 5-IAF to the mixture. The derivatization is conducted at room temperature in dark for 1 h, after which the mixtures are centrifuged at 9000× g for 5 min to sediment the insoluble proteins. 10 µL supernatant is diluted 100× with water and analyzed immediately by CE-LIF[2]. |
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References: [1]. Salvatore Sotgia, et al. Plasma L-Ergothioneine Measurement by High-Performance Liquid Chromatography and Capillary Electrophoresis after a Pre-Column Derivatization with 5-Iodoacetamidofluorescein (5-IAF) and Fluorescence Detection. PLoS One. 2013; 8(7): e70374. |
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| Cas No. | 63368-54-7 | SDF | |
| 别名 | 5-碘乙酰氨基荧光素; 5-Iodoacetamidofluorescein | ||
| Canonical SMILES | O=C(NC1=CC2=C(C3(C4=C(OC5=C3C=CC(O)=C5)C=C(O)C=C4)OC2=O)C=C1)CI | ||
| 分子式 | C22H14INO6 | 分子量 | 515.25 |
| 溶解度 | DMF: 30 mg/ml,DMSO: 30 mg/ml,DMSO:PBS (pH 7.2) (1:3): 0.25 mg/ml,Ethanol: 500µ g/ml | 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 | 1.9408 mL | 9.704 mL | 19.4081 mL |
| 5 mM | 0.3882 mL | 1.9408 mL | 3.8816 mL |
| 10 mM | 0.1941 mL | 0.9704 mL | 1.9408 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。
Quality Control & SDS
- View current batch:
- Purity: >95.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet