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| CPI-1189necrosis factor (TNF) alpha inhibitor |
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 = 99.28%
- COA (Certificate Of Analysis)
- HPLC
- NMR (Nuclear Magnetic Resonance)
- MSDS (Material Safety Data Sheet)
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Chemical structure
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| Cas No. | 183619-38-7 | SDF | Download SDF |
| Chemical Name | (Z)-N-(4-(tert-butylcarbamoyl)phenyl)acetimidic acid | ||
| Canonical SMILES | C/C(O)=N/C1=CC=C(C=C1)C(NC(C)(C)C)=O | ||
| Formula | C13H18N2O2 | M.Wt | 234.29 |
| Solubility | ≥23.4mg/mL in DMSO | Storage | Store at -20°C |
| 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. | ||
CPI-1189 is a proapoptotic cytokine tumor necrosis factor (TNF) alpha inhibitor.
AIDS dementia complex is characterized by increased gliosis, apoptosis, and oxidative stress in the CNS, as well as a compromised blood-brain barrier. TNF-alpha has been reported to be elevated in AIDS dementia complex brains and may contribute to AIDS dementia complex.
In vitro: CPI-1189 could potently inhibit p38-MAPK phosphorylation displaying protective action against tumor necrosis factor-alpha (TNFalpha)-induced neurodegeneration. Moreover, in primary astrocytes treated with interleukin 1beta, CPI-1189 inhibited p38-MAPK phosphorylation at low concentrations [1].
In vivo: To model elevated TNF-alpha in AIDS dementia complex, TNF-alpha was infused into rats. Results showed that the co-administration of CPI-1189 prevented TNF-alpha induced apoptosis. Both TNF-alpha and CPI-1189 treatment could suppress glial fibrillary acidic protein staining. TNF-alpha alone did not affect the integrity of the blood-brain barrier significantly, but CPI-1189 treatment increased blood-brain barrier integrity [2].
Clinical trial: Previous clinical study showed that CPI-1189 was well tolerated, with 91% of CPI-1189-treated subjects and 76% of placebo-treated subjects completing the trial. Skin rash was equally found in placebo and drug-treated arms. One subject developed a cataract on CPI-1189. CD4 lymphocyte counts and plasma HIV viral load remained stable in all groups throughout the trial [3].
References:[1] Hensley K,Robinson KA,Pye QN,Floyd RA,Cheng I,Garland WA,Irwin I. CPI-1189 inhibits interleukin 1beta-induced p38-mitogen-activated protein kinase phosphorylation: an explanation for its neuroprotective properties Neurosci Lett.2000 Mar 10;281(2-3):179-82.[2] Bjugstad KB,Flitter WD,Garland WA,Philpot RM,Kirstein CL,Arendash GW. CPI-1189 prevents apoptosis and reduces glial fibrillary acidic protein immunostaining in a TNF-alpha infusion model for AIDS dementia complex. J Neurovirol.2000 Dec;6(6):478-91.[3] Clifford DB,McArthur JC,Schifitto G,Kieburtz K,McDermott MP,Letendre S,Cohen BA,Marder K,Ellis RJ,Marra CM;Neurologic AIDS Research Consortium. A randomized clinical trial of CPI-1189 for HIV-associated cognitive-motor impairment. Neurology.2002 Nov 26;59(10):1568-73.
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我要做的两个目的蛋白，分子量一个是140的，另一个是36，内参用的GAPDH。这几天刚刚要做小鼠组织的蛋白，就把新提取的组织蛋白和之前做过的细胞蛋白一起上了，细胞和组织的各上了3个孔。
电泳转膜孵育等条件都是以前做成熟的
之后显影，细胞蛋白的，两个目的和内参都出得挺好；但是小鼠的组织蛋白只有分子量140的那个显出条带了，36的那个显出来的像一片水渍一样，斑片状的黑，内参有时也这样、有时干脆什么都不显。不知道这是什么原因...按说细胞同样的蛋白做出来，WB的各个步骤应该不会有太大问题的。
有点怀疑蛋白提取的不好，我提组织蛋白的的操作是在冰上进行的，裂解液也加有PMSF，用BCA法测得的蛋白浓度大概也有3~5mg/ml。
在这里求助各位高手~望给我指点一下~不胜感激啊！
分子式：C14H22O(C2H4O)n
用途：Triton X-100是一种比较温和的去垢剂（表面活性剂或称界面活性剂）,常作为添加剂使蛋白保持稳定,尤其是膜蛋白.
性质：
1.Triton X-100对细菌等微生物没有杀伤作用.
2.Triton X-100在紫外波段下有光吸收（lambda max = 275 nm and 283 nm in methanol）,因此,当缓冲液中存在Triton X-100时不能通过测定280nm光吸收来进行蛋白定量.
3.Triton X-100常与CHAPS等zwitterionic 去垢剂一起使用来纯化膜蛋白,使膜蛋白保持其天然构象.
4.Triton X-100可以提高真核细胞细胞膜的通透性,因此在Immunostaining时常使用含 0.1%-0.5%Triton X-100的PBS或TBS.
5.Triton X-100非常稳定,密封避光 +2 to +8℃条件下可以长期保存.
6.Triton X-100是透明,略显粘稠,稍微发黄的液体.
7.Triton X-100的密度为1.07 g/ml,pH6.0-8.0（5%的溶液）.
8.Triton X-100可以高压灭菌.
9.用Triton X-100来裂解细胞时,0.1% Triton X-100就足够了,不过,达到0.5%时也没问题.
10.蛋白酶K在含1% Triton X-100的溶液中依然保持活性.
11.可以使用Amberlite hydrophobic XAD resins and Rezorian A161 cartridges 来去除Triton X-100 .
去垢剂(又称表面活性剂)是一类即具有亲水基又具有疏水基的物质,一般具有乳化、分散、和增溶作用,可分阴离子、阳离子和中性去垢剂等多种类型,中性去垢剂在蛋白提取中应用的较多.
中性去垢剂又称非离子表面活性剂,对蛋白质的变性作用影响较少,宜于蛋白质或酶提取之用.一般市售中性去垢剂有聚乙二醇类,如PEG200；多元醇类表面活性剂,如山梨醇、司盘类和吐温类；聚氧乙烯脂肪醇醚,如苄泽类、平平加类；聚氧乙烯烷基苯酚醚,如Igepal CO、乳化剂OP、Triton、Pluronic(用作消泡剂、润湿剂、增溶剂)、泡敌.中性去垢剂作用后可通过Sephadex LH-50柱除去；也可直接上DEAE-Sephadex柱层析分离目的蛋白,不必先除去去垢剂.
如要膜蛋白保持原来的结构应采用：非离子型去垢剂.
RAW细胞，6孔板，RIPA提取蛋白（提前加好PMAF），每孔100uLRIPA，冰上用tip倒过来刮细胞，放置15min，12000gX15min离心，取上清。做WB时，沸水煮5min,胶冻样物质还是存在。
操作过程中发现离心后还是有胶冻样物质，和上清混在一起无法分开，严重的时候基本上全是胶冻样物质。
这个胶冻样物质应该是DNA，园友提供的方法为用超声碎裂器，但是我们实验室没有。请问还有别的方法可以使用么？
