LY2606368 HClCHK1 inhibitor |
Sample solution is provided at 25 µL, 10mM.
Quality Control & MSDS
- View current batch:
- Purity = 98.00%
- COA (Certificate Of Analysis)
- MS (Mass Spectrometry)
- NMR (Nuclear Magnetic Resonance)
- MSDS (Material Safety Data Sheet)
Chemical structure
Cell experiment [1]: | |
Cell lines | Hela cells |
Preparation method | Limited solubility. 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. |
Reaction Conditions | 7 h |
Applications | LY2606368 HCl triggers DNA damage during S-phase as pH2AX (S139) and TUNEL-positive staining cells increases substantially in Sphase cells. LY2606368 HCl also need CDC25A and CDK2 to trigger DNA damage. In addition, LY2606368 HCl leads to replication catastrophe. |
Animal experiment [1]: | |
Animal models | Female CD-1 nu-/nu- mice (26–28 g) bearing Calu-6 tumor |
Dosage form | Twice daily for 3 days with 1, 3.3, or 10 mg/kg of LY2606368 HCl |
Applications | Up to 72.3% tumor growth inhibition is observed in all three doses of LY2606368 HCl groups. Wight loss of mice is not exceeded by 3%, indicating the LY2606368 HCl is well tolerated in any of the treatment groups. Moreover, tumor regrowth of the highest dose group is slow during the 28-day recovery period, suggesting a durable response to LY2606368 HCl. |
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. King C, Diaz HB, McNeely S et al. LY2606368 Causes Replication Catastrophe and Antitumor Effects through CHK1-Dependent Mechanisms. Mol Cancer Ther. 2015 Sep;14(9):2004-13. |
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Cas No. | N/A | SDF | Download SDF |
Synonyms | N/A | ||
Chemical Name | 5-((5-(2-(3-aminopropoxy)-6-methoxyphenyl)-1H-pyrazol-3-yl)amino)pyrazine-2-carbonitrile hydrochloride | ||
Canonical SMILES | NCCCOC1=CC=CC(OC)=C1C2=CC(NC(C=N3)=NC=C3C#N)=NN2.Cl | ||
Formula | C18H20ClN7O2 | M.Wt | 401.85 |
Solubility | ≥6.63mg/mL in DMSO with gentle warming | 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. |
LY2606368 HCl is a selective ATP competitive inhibitor of checkpoint kinase 1(CHK1) with IC50 value of 1.5nM in SW1990 cells [1].
CHK1 is an intracellular serine/threonine kinase that plays a role in DNA damage response pathway. The inhibitors of CHK1 are developed for the treatment of cancers. LY2606368 HCl is an ATP-competitive inhibitor of CHK1 and is undergoing clinical trials currently. It inhibits the auto-phosphorylation of CHK1 and induces the phosphorylation of H2AX in cancer cells. In the pancreatic cell line SW1990, LY2606368 HCl significantly inhibits cell proliferation with IC50 value of 1.5nM. LY2606368 HCl also exerts potent anti-tumor activity in SW1990 xenograft model. Besides that, in the orthotopic SKVO3 model, treatment of LY2606368 HCl is found to inhibit tumor growth and reduce the incidence of metastases and accumulation. However, LY2606368 HCl is only administered intravenously due to its poor oral bioavailability [1, 2 and 3].
References:[1] Wu W, Bi C, Bence A K, et al. Antitumor activity of Chk1 inhibitor LY2606368 as a single agent in SW1990 human pancreas orthotopic tumor model. Cancer Research, 2012, 72(8 Supplement): 1776.[2] Lainchbury M, Matthews T P, McHardy T, et al. Discovery of 3-alkoxyamino-5-(pyridin-2-ylamino) pyrazine-2-carbonitriles as selective, orally bioavailable CHK1 inhibitors. Journal of medicinal chemistry, 2012, 55(22): 10229-10240.[3] McNeely S C, Burke T F, DurlandBusbice S, et al. Abstract A108: LY2606368, a second generation Chk1 inhibitor, inhibits growth of ovarian carcinoma xenografts either as monotherapy or in combination with standard-of-care agents. Molecular Cancer Therapeutics, 2011, 10(Supplement 1): A108.
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protocol如下:
灌注取脑后,4%多聚甲醛4℃固定24-48h。修块3-4mm。
脱水(常温):45%、55%、65%、75%、85%、95%、100%酒精各30min;
透明(常温):(100%酒精+二甲苯)/210min,二甲苯①20min,二甲苯②至完全透明(约20min)
浸蜡(60℃):(二甲苯+石蜡①)/210min,石蜡①30min,石蜡②40min;石蜡①为低熔点,石蜡②为高熔点
包埋:浸蜡后用石蜡②手动包埋;
切片厚度:6um、(8um也切的有)
展片:40℃水中展片;
烤片:60摄氏度烤片机侧烤约5min后吸去水分,转移至60℃温箱烤约4h;
脱蜡:烤片后趁蜡未凝固,进行脱蜡。二甲苯5min×2,100%酒精3min,95%酒精2min,
85%酒精2min,75%酒精2min,流水冲洗2min(放在缸子里用水泡的,换了几次水);
染色:
HE染色:苏木精5-8min(新配为5min,时间长可为8min,用前过滤),流水冲洗3min
1%盐酸酒精分化5s,流水冲洗1min,0.1%伊红1-2min,
脱水:85%酒精2min,95%酒精2min,100%酒精2min,
100%无水乙醇:二甲苯(1:1)2min
透明:二甲苯(I)3min,二甲苯(II)3min;
CV染色:浸入CV染液中10-20min,
脱水:75%乙醇2min,85%乙醇2min,95%乙醇2min,100%无水乙醇2min,
100%无水乙醇:二甲苯(1:1)2min
透明:二甲苯(I)3min,二甲苯(II)3min;
封片:中性树胶封片(整个染色过程避免干片)。
部分HE染色、CV染色片子结果如图所示,基本上没有完整的片子(染色时,片子随着在液体里的时间脱片、烂片的程度加深),另CV染色着色浅。
请教战友是什么原因?哪里出了问题,该怎么解决?
尤其是前面两个,后面的还可以从字面理解着做,前面这两个就纯把我搞晕菜了.
全量法:测水浸出物的
酒石酸铁比色法;
紫外分光光度法;
恒重法:测水含量的
谢谢谢谢
红色的是胰岛素,绿色的是胰高血糖素。都是石蜡切片~
请问下我这染出来的应该是胰岛细胞吧,背景颜色适合吗?
为什么胰高血糖素然不出来,这两种我用的方法是一样的。
方法:
1.脱蜡,水化:脱蜡前应将组织芯片在60摄氏恒温箱中烘烤1h
A:组织芯片置于二甲苯中浸泡15min,更换二甲苯后再浸泡10min(脱蜡)
B:无水乙醇中浸泡5min(水化)
C:95%乙醇中浸泡5min
D:75%乙醇中浸泡5min
2.PBS洗2次各5min
3.PBS洗2次每次5min,洗去枸橼酸钠(Ⅲ,Ⅱ)
4.3%H2O2(80%甲醇稀释)室温静置10min(去除内源性酶)
5.PBS洗2次各5min(Ⅰ、Ⅱ)
6.加入1%BSA稀释(0.01MPBS稀释)封闭液,37度恒温烘箱1h,顷去多余液体,不洗
7.滴加一抗用1%BSA稀释(1:50)或(1:100),先室温放置1h,然后4度过夜(放入湿盒)并用PBS代替一抗做阴性对照
8.PBS洗3次每次5min
9.滴加二抗(1:100)或(1:200),用1%BSA稀释37度恒温箱(放入湿盒)1h,避光
10.PBS洗3次各2min,避光,用摇床洗涤
11.加入20ul样品Hoechst液体(10ug/ml)5min用摇床避光
12.PBS洗3次各2min,避光,用摇床洗涤
13.滴加抗荧光猝灭剂,封片,镜检,绿色荧光
主药:50mg
mcc:20mg
l-hpc:10mg
cms-na:10mg
5%pvp50%乙醇qs
外加
ms:0.5mg
cms-na:3mg
50度干燥
以上处方不粘冲,但因上处方的辅料与主药有相互作用,导致片子变黄
现换成以下处方
处方1
主药:50mg
预胶化淀粉:20mg
淀粉:20mg
8%淀粉浆:qs
外加
ms:0.5或1mg
滑石粉:3或5mg
60度干燥
以上处方粘冲,为何?
处方2
主药:50mg
预胶化淀粉:20mg
淀粉:10mg
甘露醇:10mg
8%淀粉浆:qs
外加
ms:0.5或1mg
滑石粉:3或5mg
60度干燥
以上处方粘冲,为何
[国家标准物质网]高频红外碳硫分析仪是一种常用的分析仪器,能快速、准确地测定钢、铁、合金、有色金属、水泥、矿石、玻璃及其它材料中碳、硫两元素的质量分数。高频红外碳硫分析仪在使用过程中需要进行定期维护,今天我们就来具体介绍一下高频红外碳硫分析仪的日常维护方法。
高频红外碳硫分析仪日常维护要点
1、高频红外碳硫分析仪分析样品所用的坩埚需要马弗炉中加热温度升到1000度到1200度,恒温2小时后,自然冷却至一定温度时,取出放干燥器中冷至室温备用。
2、分析低含量样品所用的钨粒需在恒温箱中,加热至200度烘2小时后,取出在干燥器中冷却后使用。
3、高频红外碳硫分析仪设置了高频输出功率调节,在未打开仪器电源之前,用万用表测试电网电压后,将换档开关拨至电网电压基本一致的档位上,开始调试仪器。方法是当然界电网电压为220V时,调至230V档为减小输出功率,调至210档为增加输出功率。
4、打开总氧气阀门,打开仪器电源,按技术员所教操作。
5、每天仪器示打开电源前或分析200个样品后,需清扫一次过滤网及整个炉腔。
6、每天应检查高频炉正前方的干燥剂和脱脂棉。一旦发现干燥剂1/3变红或结块,都应立即更换。脱脂棉长期使用1/3变黄后,也应立即更换。更换时将旋转试管下方的螺杆,使试管下降至能离开上端面为止,取出试管,更换后按原装好拧紧,检查不漏气为止。
7、每月应清洗过滤网一次,清洗方法是:按上述“清扫方法”取出过滤网后,再取出过滤网上、下两只密封圈,将粉尘清扫后,放入超声波清洗器中,在清洗槽内加入蒸馏水和洗洁精。打开超声波清洗器电源。30分钟后取出过滤网,用蒸馏水反复清洗后,用无水乙醇清洗。最后取出过滤网用电吹风干或放入烘箱烘干,装上、下O型密封圈,涂上真空硅脂,按原位老妈子,检查不漏气为止。
8、高频红外碳硫分析仪使用时每季度应检查经过压紧阀内部的硅橡胶管,若发现有老化或弹性不好时应立即更换。
摘自:国家标准物质网