潮霉素B；Hygromycin B
分子式：C20H37N3O13 分子量：527.52
物理性状及指标：
外观：……………………Off-white powder
溶解性：…………………50mg/ml in water
使用方法推荐（仅供参考）
储运液配置：溶解于无菌细胞培养级1M HEPES buffer (MA0036)或者PBS1X(MA0015)，配置成浓度为50mg/ml. 无菌微膜过滤。然后根据自身实验要求进行稀释或者使用
筛选条件：Most cells growing aerobically are killed by Hygromycin B in the concentration range of 50 to 500 μg/ml. However, the sensitivity of cells is pH dependent (i.e. the higher the pH of the culture medium the greater the sensitivity). Thus, the concentration of Hygromycin B required for complete growth inhibition of given cells can be reduced by increasing the pH of the medium. In addition, you can also lower the required amount of Hygromycin B Gold by using low‑salt media whenever possible.
Escherichia coli (大肠杆菌)
Hygromycin-resistant transformants are selected in low‑salt LB agar medium (yeast extract 5g/l, tryptone 10 g/l, NaCl 5 g/l, agar 15 g/l, pH 8) supplemented with 50 to 100 μg/ml of Hygromycin B. Plates containing Hygromycin B are stable for 1 month when stored at 4 °C.
Mammalian cells （哺乳动物细胞）
The working concentrations of Hygromycin B for mammalian cell lines vary from 50 to 200 μg/ml; in a few cases, up to 500 μg/ml. In a starting experiment we recommend to determine the optimal concentration of Hygromycin B required to kill your host cell line. Killing and detachment of dead cells from the plate, especially at high cell density, can require a longer time than with G418. Hygromycin‑resistant stable transfectants are usually obtained after 10 days to 3 weeks incubation, depending on the cell line. See table on the next page for suggested working concentrations of Hygromycin B in mammalian cells.
工作浓度
Hygromycin B normally used at a concentration of 200 μg/ml, a 500‑fold dilution from the stock solution. However, the optimal concentration needs to be determined for your cells. Suggested concentrations of Hygromycin B for selection in some examples of mammalian cells are listed below.
| Cell line |
Medium |
Hygromycin B conc. |
|
B16 (Mouse melanocytes) |
RPMI |
100-200 μg/ml |
|
CHO (Chinese hamster ovarian cells) |
MEM |
100-500 μg/ml |
|
HeLa (Human uterine cells) |
DMEM |
100-200 μg/ml |
|
HEK293 (Human embryonic kidney cells) |
DMEM |
50-400 μg/ml |
|
Raji (Human lymphocytes) |
RPMI |
125-300 μg/ml |
|
THP-1 (Human monocytes) |
RPMI |
250-400 μg/ml |
用途及描述：科研试剂，阻止多肽合成和抑制延长。用于原核和真核细胞的选择和保藏。潮霉素B（Hygromycin B）是由吸水链霉菌（Streptomyces hygroscopicus）代谢产生的一种氨基糖苷类抗生素，通过干扰70S核糖体易位和诱导对mRNA模板的错读而抑制蛋白合成，从而杀死原核（如细菌）、真核（如酵母菌，真菌）和高等哺乳动物真核细胞。
储存条件：2-8℃，避光防潮密闭干燥。
运输条件：常温运输
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|
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|
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|
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英文名字：Hygromycin B
≥90%,粉末,进分
|
TESTS |
SPECIFICATIONS |
Appearance |
Off-white powder |
Solubility |
50mg/ml in water |
Identification |
HNMR |
Specific rotation |
+18.0 to +23.0° |
Activity |
≥950U/mg |
Purity |
≥90% |