Phleomycin
(Synonyms: 腐草霉素,Zeocin) 目录号 : GC14081
Phleomycin是一种糖肽类抗生素，是一种DNA链断裂（DNA-SB）诱导剂，可用于筛选基因工程细胞。
Cas No.:11006-33-0
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
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| Cell experiment [1]: | |
Cell lines | HeLa cells |
Preparation Method | Cells (6×104 per well) were allowed to attach in 24-well plates for 1 day before adding Phleomycin and cultures were measured at 0, 6 , 12, 18, 24h and 100, 200, 300, 400μg/mL of Phleomycin with a hemacytometer. The cytotoxic effects of Phleomycin on HeLa cells was determined by a viable cell count and MTT assay. |
Reaction Conditions | 100, 200, 300, 400μg/mL; 0-24h |
Applications | Phleomycin inhibits HeLa cells proliferation and induces cytotoxicity in a dose-dependent and time-dependent manner. |
References: | |
Phleomycin is a glycopeptide antibiotic and a DNA strand break (DNA-SB) inducer that can be used to select genetically-engineered cells[1]. Phleomycin is a molecule found in Streptomyces, and similar to bleomycin, can bind to and insert into DNA, disrupting the integrity of the double helix structure[2]. Phleomycin has a mutagenic effect on resistant Streptococcus strains[3].
In vitro, treatment of HeLa cells with Phleomycin (100-400μg/mL) for 0-24h inhibited cell proliferation in a time- and dose-dependent manner, induced cytotoxicity, induced the initial release of cytochrome c, downregulation of Bcl-XL, ENDOG, DAXX, and MDM2, and upregulation of CASP and BID[4]. Treatment of Nannochloropsis oceanica cells with Phleomycin (0.2μg/mL) for 48h caused random double-strand breaks in the DNA of the algal cells and increased the transformation efficiency of the active hygromycin B resistance gene (aph7)[5]. Phleomycin (0-200ng/mL) treatment of mouse embryonic fibroblasts (MEFs) from different mutants for 48h significantly reduced the survival rate of MEFs lacking XRCC4, DNA-Ligase IV or Xlf, but had no significant effect on MEFs from PAXX mutants and wild-type MEFs[6].
References:
[1] Ma X, Pan K, Zhang L, et al. Genetic transformation of Nannochloropsis oculata with a bacterial phleomycin resistance gene as dominant selective marker[J]. Journal of Ocean University of China, 2016, 15: 351-356.
[2] Stokowa-Sołtys K, Dzyhovskyi V, Wieczorek R, et al. Phleomycin complex–Coordination mode and in vitro cleavage of DNA[J]. Journal of inorganic biochemistry, 2019, 195: 71-82.
[3] Van Peer A F, De Bekker C, Vinck A, et al. Phleomycin increases transformation efficiency and promotes single integrations in Schizophyllum commune[J]. Applied and Environmental Microbiology, 2009, 75(5): 1243-1247.
[4] Hwang J, Kim Y Y, Huh S, et al. The Time‐Dependent Serial Gene Response to Zeocin Treatment Involves Caspase‐Dependent Apoptosis in HeLa Cells[J]. Microbiology and immunology, 2005, 49(4): 331-342.
[5] Zhang Z, Guo L, Liu H, et al. Zeocin treatment significantly elevated transformation efficiency of Nannochloropsis oceanica[J]. Journal of Applied Phycology, 2022, 34(3): 1587-1594.
[6] Abramowski V, Etienne O, Elsaid R, et al. PAXX and Xlf interplay revealed by impaired CNS development and immunodeficiency of double KO mice[J]. Cell Death & Differentiation, 2018, 25(2): 444-452.
Phleomycin是一种糖肽类抗生素，是一种DNA链断裂（DNA-SB）诱导剂，可用于筛选基因工程细胞[1]。Phleomycin是在链霉菌中发现的，与博来霉素 （Bleomycin）类似，它能够结合并插入DNA，破坏双螺旋结构的完整性[2]。Phleomycin对抗性链球菌菌株具有诱变作用[3]。
在体外，Phleomycin（100-400μg/mL）处理HeLa细胞0-24h，以时间和剂量依赖性方式抑制了细胞增殖，诱导了细胞毒性，诱导了细胞色素c的初始释放、Bcl-XL、ENDOG、DAXX和MDM2的下调以及CASP和BID的上调[4]。Phleomycin（0.2μg/mL）处理微拟球藻（Nannochloropsis oceanica）细胞48h，引起了藻细胞DNA的随机双链断裂，提高了活性潮霉素B抗性基因（aph7）的转化效率[5]。Phleomycin（0-200ng/mL）处理来自不同突变体的鼠胚胎成纤维细胞（MEFs）48h，显著降低了缺乏XRCC4、DNA-Ligase IV或Xlf的MEFs的存活率，但对来自PAXX突变体的MEFs和野生型MEFs无显著影响[6]。
| Cas No. | 11006-33-0 | SDF | |
| 别名 | 腐草霉素,Zeocin | ||
| 化学名 | (2R,3S,4S,5R,6R)-2-(((2R,3S,4S,5S,6S)-2-(2-(6-amino-2-(3-amino-1-((2,3-diamino-3-oxopropyl)amino)-3-oxopropyl)-5-methylpyrimidine-4-carboxamido)-3-((5-((1-((2-(4-((4-guanidinobutyl)carbamoyl)-4',5'-dihydro-[2,4'-bithiazol]-2'-yl)ethyl)amino)-3-hydroxy-1-o | ||
| Canonical SMILES | NC(CNC(CC(N)=O)C1=NC(N)=C(C(C(NC(C(NC(C(C(C)C(NC(C(NCCC2=NC(C3=NC(C(NCCCCNC(N)=N)=O)=CS3)CS2)=O)C(O)C)=O)O)C)=O)C(O[C@H]4[C@]([H])([C@H]([C@@H]([C@@H](O4)CO)O)O)O[C@H]5O[C@@H]([C@H]([C@@H]([C@@H]5O)OC(N)=O)O)CO)C6=CNC=N6)=O)=N1)C)C(N)=O.[Cu+2].Cl | ||
| 分子式 | C55H85N20O21S2Cu • HCl | 分子量 | 1526.5 |
| 溶解度 | ≤10mg/ml in PBS, pH 7.2 | 储存条件 | Store at 2-8°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 | 0.6551 mL | 3.2755 mL | 6.5509 mL |
| 5 mM | 0.131 mL | 0.6551 mL | 1.3102 mL |
| 10 mM | 0.0655 mL | 0.3275 mL | 0.6551 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。