| Misonidazolenitroimidazole with radiosensitizing and antineoplastic properties |

Sample solution is provided at 25 µL, 10mM.
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Cell Stem Cell.2017 Nov 20. pii: S1934-5909(17)30375-2.Quality Control & MSDS
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| Cas No. | 13551-87-6 | SDF | Download SDF |
| Synonyms | NSC 261037,Ro 7-0582,SR 1354 | ||
| Chemical Name | α-(methoxymethyl)-2-nitro-1H-imidazole-1-ethanol | ||
| Canonical SMILES | OC(COC)CN1C=CN=C1[N+]([O-])=O | ||
| Formula | C7H11N3O4 | M.Wt | 201.2 |
| Solubility | ≤5mg/ml in ethanol;15mg/ml in DMSO;15mg/ml in dimethyl formamide | Storage | Store at -20°C |
| Physical Appearance | A crystalline solid | Shipping Condition | Evaluation sample solution : ship with blue ice.All other available size:ship with RT , or blue ice upon request |
| General tips | For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months. | ||
Misonidazole (MISO), a hypoxic cell radiosensitizer, is selectively metabolized by hypoxic cells to reactive products that bind covalently to cellular constituents. The drug also possesses a substantial cytotoxic effect, independent of radiation, which is selectively expressed in hypoxic cells. Misonidazole may be cytotoxic to the normal hypoxic tissues in the human body, making this became a major concern in the clinical application of the drug. Misonidazole leads to strand breaks in cellular DNA and those cells which fail to survive also fail to repair these strand breaks. Misonidazole depletes intracellular glutathione and is more toxic in glutathione depleted cells. The depletion is time, temperature, drug concentration and cell line dependent.
In vitro: In cultured CHO cells, pretreatment with 5 mM MISO for 2 hr exihibited the marginal toxicity [2].
In vivo: Pretreatment with misonidazole (MISO) enhanced the number of cross-links formed in a fibrosarcoma and in the spleen and gut of mice for periods up to 48 h following a single injection of melphalan (MEL). MISO pretreatment could result in a greater amount of binding of MEL to DNA at early times after injection. MISO may exert its affect by inhibiting the repair of cross-links or monoadducts at early times post-injection [3].
Clinical trials: Various dose schedules of misonidazole have proven to be tolerable, with a moderate incidence of nausea and vomiting and mild peripheral neuropathy, and a low incidence of more severe peripheral neuropathy or central neuropathy [4].
References:[1] Josephy P D, Palcic B, Skarsgard L D. Ascorbate-enhanced cytotoxicity of misonidazole[J]. Nature, 1978, 271(5643): 370-372.[2]Taylor Y C, Bump E A, Brown J M. Studies on the mechanism of chemosensitization by misonidazole in vitro[J]. International Journal of Radiation Oncology Biology Physics, 1982, 8(3-4): 705-708.[3] Murray D, Meyn R E. Enhancement of the DNA cross-linking activity of melphalan by misonidazole in vivo[J]. British journal of cancer, 1983, 47(2): 195.[4] Wasserman T H, Stetz J A, Phillips T L. Clinical trials of misonidazole in the United States[J]. Cancer clinical trials, 1980, 4(1): 7-16.
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利用NheI和HindIII双酶切插入目的片段,为什么双酶切有目的片段和切开的质粒,测序却没有?
并且测序后在NheI位点后的序列变成了-GGCTAGGTAC-Kpn位点(CGTTTAAACTTAAGCTTG消失。之前和之后的序列都相符),之间的怎么都没了?
请教高人,这样的问题怎么解决?
多谢
你所说的“测序酶”在一般情况下就是“聚合酶”。
因为目前的的测序方法主要就是借助聚合反应。
当然,有些测序方法(比如曾经的SOLiD系统)是利用连接反应,那么它用的“测序酶”肯定是连接酶。
估计,也就是这个原因有人把它。
两个基因大小分别230kb和450kb,酶切后,450kb的有目的条带,但很弱,230kb的质粒条带亮,其下方有一很微弱条带,用的酶分别是:Ncol和Spel体系是(Takara,两个酶切体系不同,用的官网推荐体系):
NcoI1μl
Spel1μl
10×KBuffer2μl
0.1%BSA2μl
DNA3ul
灭菌水upto20μl37℃4h电泳1h
期待着高手们的指点,谢谢。
【之前构建的4个重组质粒里也有两个出现这样的问题(aagtcc变成agtcc),但是酶切能切出插入的片段】
目前问测序公司得到的答复是样品可能不是单克隆,测序结果是不准确的,建议重新挑取单克隆测序。
不知道还有没有别的方法?
●应用酶水解多肽不会破坏氨基酸,也不会发生消旋化。水解的产物为较小的肽段。向左转|向右转

