Hygromycin B
(Synonyms: 潮霉素; 潮霉素B; Hygrovetine) 目录号 : GC15496
Hygromycin B 是由吸水链霉菌产生的氨基糖苷类抗生素。
Cas No.:31282-04-9
Sample solution is provided at 25 µL, 10mM.
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Hygromycin B is an aminoglycoside antibiotic produced by Streptomyces hygroscopicus. Widely used in veterinary medicine and in cell culture selections, it kills bacteria, fungi, and higher eukaryotic cells, including mammalian cells [1].
The sensitivity of various cultured cell lines to Hygromycin B was assessed by plating cells at low density in Dulbecco modified Eagle medium supplemented with 10% fetal calf serum and Hygromycin B to give final drug concentrations ranging between 50 and 400 ug/ml of medium. Usually one or two cycles of replication still occurred before the onset of cytotoxicity; cell death commenced ca.3 days after the beginning of drug treatment and was generally complete after 8days (CV1 and HeLa cells sometimes required 10 to 12 days before cell killing was complete). Sofar, no cellline has been found that is naturally resistant to Hygromycin B [2].
The E.coli bacterial Hygromycin B resistance gene provides a basis for testing the usefulness of Hygromycin B as adominant selectable marker in Hygromycin B -susceptible cells. It may also be possible to use promoter less coding sequences of the Hygromycin B resistance gene as a promoter probe [3].
Hygromycin B provides a generally applicable selection system for DNA transfer experiments between both procaryotic and eukaryotic cells [2].
References:
[1]. Borovinskaya MA, Shoji S, Fredrick K, Cate JH. 2008. Structural basis for hygromycin B inhibition of protein biosynthesis. RNA 14: 1590–99
[2]. KAREN BLOCHLINGER，HEIDI DIGGELMANN. Hygromycin B Phosphotransferase as a Selectable Markerfor DNA Transfer Experiments with Higher Eucaryotic Cells. MOLECULAR AND CELLULAR BIOLOGY, Dec.1984, p.2929-2931
[3]. R. N. Rao, N. E. Allen, J. N. J. Hobbs, W. E. J. Alborn, H. A. Kirst & J. W. Paschal: Genetic and enzymatic basis of hygromycin B resistance in Escherichia coli. Antimicrob. Agents Chemother. 24, 689-695 (1983)
Hygromycin B 是由吸水链霉菌产生的氨基糖苷类抗生素。广泛用于兽医和细胞培养选择，可杀死细菌、真菌和高等真核细胞，包括哺乳动物细胞[1]。
通过在添加了 10% 胎牛血清和潮霉素 B 的 Dulbecco 改良 Eagle 培养基中以低密度铺板细胞，评估各种培养细胞系对潮霉素 B 的敏感性，最终药物浓度范围为 50 至 400 微克/毫升中等的。通常在细胞毒性发生之前仍会发生一到两个复制周期；细胞死亡在药物治疗开始后约 3 天开始，通常在 8 天后完成（CV1 和 HeLa 细胞有时需要 10 至 12 天才能完成细胞杀死）。目前尚未发现对潮霉素B具有天然抗性的细胞系[2]。
大肠杆菌细菌潮霉素 B 抗性基因为测试潮霉素 B 作为潮霉素 B 易感细胞中显性选择标记的有效性提供了基础。也可以使用潮霉素 B 抗性基因的无启动子编码序列作为启动子探针[3]。
潮霉素 B 为原核和真核细胞之间的 DNA 转移实验提供了一个普遍适用的选择系统[2]。
| Cell experiment [1]: | |
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Cell lines |
Wild type haploid Saccharomyces cerevisiae strain Y166 and the spontaneous CRY6 mutant |
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Preparation Method |
Logarithmically growing yeast cells (about 2- 107/ml) were converted into spheroplasts by treatment with glusulase. The spheroplasts were recovered by incubation at 30℃ for 90 min in YEPD medium plus I M sorbitol. Then the culture was divided in three aliquots and each received 10 µCi/ml of [3H]leucine (54 Ci/mmol). One aliquot served as control, the other two received 250 and 500 µg/ml hygromycin B, respectively, and incubation was continued at 30℃. |
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Reaction Conditions |
50 and 500 µg/ml,30℃,20-120 min. |
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Applications |
Protein synthesis by yeast spheroplasts is blocked by hygromycin B as determined by the uptake of [3H]leucine into trichloroacetic acid-precipitable material |
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References: [1]: Gonzalez A, Jimenez A, Vazquez D, Davies JE, Schindler D. Studies on the mode of action of hygromycin B, an inhibitor of translocation in eukaryotes. Biochim Biophys Acta 1978; 521:459-469. |
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| Cas No. | 31282-04-9 | SDF | |
| 别名 | 潮霉素; 潮霉素B; Hygrovetine | ||
| 化学名 | (3'R,3aS,4S,4'R,5'R,6R,6'R,7S,7aS)-4-[(1R,2S,3R,5S,6R)-3-amino-2,6-dihydroxy-5-(methylamino)cyclohexyl]oxy-6'-[(1S)-1-amino-2-hydroxyethyl]-6-(hydroxymethyl)spiro[4,6,7,7a-tetrahydro-3aH-[1,3]dioxolo[4,5-c]pyran-2,2'-oxane]-3',4',5',7-tetrol | ||
| Canonical SMILES | CNC1CC(C(C(C1O)OC2C3C(C(C(O2)CO)O)OC4(O3)C(C(C(C(O4)C(CO)N)O)O)O)O)N | ||
| 分子式 | C20H37N3O13 | 分子量 | 527.5 |
| 溶解度 | ≥ 26.375mg/mL in Water | 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 | 1.8957 mL | 9.4787 mL | 18.9573 mL |
| 5 mM | 0.3791 mL | 1.8957 mL | 3.7915 mL |
| 10 mM | 0.1896 mL | 0.9479 mL | 1.8957 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: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet