- Cesium chloride
- DAPK Substrate Peptide
- GRI 977143
- Mdivi 1
- NQDI 1
| MelphalanDNA alkylating agent |
Sample solution is provided at 25 µL, 10mM.
Nature.2017 Jan 19;541(7637):417-420.
Nature.2018 Nov;563(7731):407-411.Nature.2018 Jun 13.Nature.2018 Jun 27.Nature.2018 Mar 29;555(7698):673-677.Nature.2017 Sep 7;549(7670):96-100.Nature.2016 Apr 21;532(7599):398-401.
Science.2016 Aug 5;353(6299)594-8Nat Nanotechnol.2017 Dec;12(12):1190-1198.Nature Biotechnology.2017 Jun;35(6):569-576Nat Med.2018 Sep 17.Cell.2018 Dec 21. pii: S0092-8674(18)31561-7.Cell.Available online 25 October 2018.Cell.2018 Sep 27. pii: S0092-8674(18)31183-8.Cell.2018 Jun 28;174(1):172-186.e21.Cell.2018 Feb 22;172(5):1007-1021.e17.Cell.2017 Nov 30;171(6):1284-1300.e21.Cell.2017 Aug 17. pii: S0092-8674(17)30869-3.Cell.2017 Jul 13;170(2):312-323Nat Med.2018 Jan 29.Nat Med.2017 Nov;23(11):1342-1351.Cell.2017 Apr 6;169(2):286-300.Cell.2015 Aug 27;162(5):987-1002.Cell.2015 Feb 12;160(4):729-44.Nature Medicine.2017 Apr;23(4):493-500.Cancer Cell.2018 May 14;33(5):905-921.e5.Cancer Cell.2018 Apr 9;33(4):752-769.e8.Cancer Cell.2018 Mar 12;33(3):401-416.e8.Cancer Cell.2017 Aug 14;32(2):253-267.e5.Nat Methods.2018 Jul;15(7):523-526.Cell Stem Cell.2018 May 3;22(5):769-778.e4.Cell Stem Cell.2017 Nov 20. pii: S1934-5909(17)30375-2.Quality Control & MSDS
- View current batch:
- Purity = 98.00%
- COA (Certificate Of Analysis)
- MS (Mass Spectrometry)(Retest)
- NMR (Nuclear Magnetic Resonance)
- MSDS (Material Safety Data Sheet)
- Datasheet
Chemical structure
| Cell experiment [1]: | |
Cell lines | Human myeloma cell line RPMI 8226, neuroblastoma cell lines; SH-SY5Y, SK-N-AS, and SK-N-BE |
Preparation method | The solubility of this compound in DMSO is >6.9 mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months. |
Reacting condition | 30-min |
Applications | Melphalan (5 and 10 pmol/l) resulted in an out-growth delay during the first 48 post-treatment. Melphalan treatment disclosed a relative increase of cells in S- and G-phases at 24 h followed by an accumulation of cells in G,-phase at 48 h. Treatment with high melphalan concentrations (20 and 40 pmol/l) the accumulation in the G-phase was more persistent. Melphalan treatment (20 pmol/l) dramatically decreased late S- and G,-phases. Exposure of a myeloma cell line (RPMI 8226) to a 30-minute pulse of melphalan (1-phenylalanine-mustard) resulted in a cell cycle progression delay characteristic for DNA cross-linking agents. Melphalan bound to DNA, RNA, and protein in cells in vitro. Melphalan induced chromosomal aberrations, sister chromatid exchange, micronuclei, mutations at the HPRT gene, and DNA damage in human cells in vitro. Melphalan induced transformation of C3H 10T1/2 and other cells. In cultured rodent cells, it induced chromosomal aberrations, sister chromatid exchange, gene mutations, and DNA damage. Melphalan induced aneuploidy and sex-linked recessive lethal mutations in Drosophila, and mutation in bacteria. |
| Animal experiment [3]: | |
Animal models | Immunodeficient mice bearing human ovarian tumors from A2780 cells |
Dosage form | Intraperitoneal injection, 11.7 mg/kg |
Application | In immunodeficient mice bearing human ovarian tumors from A2780 cells, melphalan (11.7 mg/kg, i.p.) severely inhibited the growth of previously untreated tumors, whereas the growth of tumors which had received prior treatment with melphalan was unaffected by the subsequent high dose. |
Other notes | Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal. |
References: [1]. Fernberg J O, Lewensohn R, Skog S. Cell cycle arrest and DNA damage after melphalan treatment of the human myeloma cell line RPMI 8226[J]. European journal of haematology, 1991, 47(3): 161-167. [2]. MELPHALAN. Pharmaceuticals.Bookshelf [3]. Caffrey P B, Zhang Y, Frenkel G D. Rapid development of drug resistance in human ovarian tumor xenografts after a single treatment with melphalan in Vivo[J]. Anticancer research, 1998, 18(4C): 3021-3025. | |
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| Cas No. | 148-82-3 | SDF | Download SDF |
| Synonyms | N/A | ||
| Chemical Name | (2S)-2-amino-3-[4-[bis(2-chloroethyl)amino]phenyl]propanoic acid | ||
| Canonical SMILES | C1=CC(=CC=C1CC(C(=O)O)N)N(CCCl)CCCl | ||
| Formula | C13H18Cl2N2O2 | M.Wt | 305.2 |
| Solubility | ≥6.85mg/mL in DMSO | Storage | Store at RT |
| Physical Appearance | A 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. | ||
Melphalan is a DNA alkylating agent and inhibits DNA and RNA synthesis [1].
DNA alkylating agent attaches the alkyl group to the guanine base of DNA and inhibits DNA and RNA synthesis, which is necessary for cells to survive.
In PC-3 cells, melphalan inhibited cells growth with IC50 values of 0.074 μg/ml and 0.77 μg/ml for sequential dosing and single dosing, respectively, which suggested the sequential dosing was more effective [1].
In 12 patients with androgen-independent prostate cancer, melphalan provided some clinical benefits with manageable toxicity and the median survival was 23 weeks [1]. In 381 myeloma patients who received melphalan-based autologous stem cell transplant (Mel-ASCT), melphalan (200 mg/m2 body surface area (BSA)) led to oral mucositis (OM) in 75% of patients. And OM was severe in 21% patients. Patients with renal dysfunction have the greatest risk for severe OM when received a high mg/kg melphalan dose [2].
References:[1]. Mougenot P, Bressolle F, Culine S, et al. In vitro cytotoxic effect of melphalan and pilot phase II study in hormone-refractory prostate cancer. Anticancer Res, 2006, 26(3B): 2197-2203.[2]. Grazziutti ML, Dong L, Miceli MH, et al. Oral mucositis in myeloma patients undergoing melphalan-based autologous stem cell transplantation: incidence, risk factors and a severity predictive model. Bone Marrow Transplant, 2006, 38(7): 501-506.
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这样可不可以？
另还有几个问题：1、这样的裂解液裂解线粒体提蛋白可不可以
2、裂解液加多少剂量
3、12000g离心10min和30分钟有没有区别
(1)1×107细胞重悬,其来源可为新鲜样品或贮存于-80℃1mol冰冻清洗缓冲液中
的无DMSO的样品,冰冻缓冲液包括10mMHEPES(pH7.5),1.5mMMgCl2,10mMKCl,
1mM二硫苏糖醇(DTT),然后离心(16000rpm、4℃、1min)。
(2)重悬细胞团于100ul冰冻裂解缓冲液,其包含10mMTris-HCl(pH7.5),1mMMgCl2,1mMEGTA,0.1mM苯甲基磺酰氟(DMSF),5mMB2巯基乙醇,0.5%CHAPs(Pierce)和10%甘油。
(3)冰育30分,然后在超速离心机中离心30分钟。
(4)去上清,液氮速冻沉淀,裂解细胞于-80℃保存。
二、扩增
(1)在微量管中冻干0.1ug荧光标记的cx反转引物,蜡封(5′CCCTTACCCTTACCC
TTACCCTTA3′)。
(2)于蜡界之上加50ulTRAP反应物,包括Tris-Cl(pH8.3)20mM,1.5mMMgCl2,
63mMKCl,0.005%,TWeen20,1mMEGTA,50mM三磷酸脱氧核苷,0.1ug荧光标记
TS前导引物(5′AATCCGTCGAGCAGAGTT3′),1ugT4基因32蛋白,牛血清白蛋白0.1mg/ml,2UTaqNDA聚合酶(Takara,shyzou)和5ugCHAPs细胞提取液。荧光标记TS引物可由市场购得。扩增前5ug提取物与1ugRNA酶37℃水浴20分钟,为了端粒酶活性的标准化,运用体内端粒酶活性标准(TTAS):TS重叠引物和CX重叠引物(TS:5′AATCCGTCGAGCAGAGTTGTGAATGAGGCCTTC3′及CX:5′CCCTTACCCTTACCCTTACCCTTATAGGCGCTCAATGTA3′)TS(18bp)和CX(24bp)序列为普通型,下划线标出的为成肌素序列(每个15bp)。成肌素CDNA通过这些引物扩
增产生了150bp的产物,它可用与ITAS扩增端粒酶梯度相同的TS引物和CX引物进行
再扩增。每一项测定用25attogram以显现I-TAS,它并不干扰TRAP测量。
(3)置于22℃10分钟,作为TS引物由端粒酶介导的延伸。90℃热灭活反应物90秒。
(4)PCR扩增,条件94℃30S,50℃30S,72℃1.5min;共27循环。
三、分析
(1)制备8%的变性胶(longRanger,ATBiochem)含6M尿素。
(2)加电泳缓冲液(1×TBE)于电泳槽。
(3)上样缓冲液与产物各1ul混合后上样,上样缓冲液含90%甲酰胺和10%蓝葡聚
糖(Bluedertian)。
(4)95℃预发性4分钟,速冷,上样,运用100bp,150bp,200bp大小荧光标记物。
(5)测序
(6)测序结果由FiagmuntManager程序自动分析,每一荧光峰根据大小、高度、面积定
量。因为过量扩增的产物会产生不可信的面积值,一部分PCR产物应在蒸馏水稀释后再分析
裂解液裂解是一种比较温和的红细胞去除方法,主要用于经酶消化分散的组织细胞的分离纯化，淋巴细胞的分离纯化以及组织细胞蛋白与核酸提取等实验中红细胞的去除。经红细胞裂解液裂解得到的组织细胞中不含红细胞，可进一步用于原代培养、细胞融合、流式细胞分析、核酸与蛋白的分离和提取等。
用SDS裂解，RNA酶降解，
在过柱，后洗脱
不知单纯使用冰浴中超声裂解够了吗？还是说要选用裂解液呢？最主要的就是我不知道该怎么赔这个裂解液，使其COX活性不产生影响
