- GSK-3 Inhibitor IX (BIO)
- SB 216763
- CHIR-99021 (CT99021)
- AZD2858
- LY2090314
| CHIR-98014GSK-3β inhibitor,selective and ATP-competitive |
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 = 99.13%
- COA (Certificate Of Analysis)
- HPLC
- NMR (Nuclear Magnetic Resonance)
- MSDS (Material Safety Data Sheet)
- Datasheet
Chemical structure
| Description | CHIR-98014 is a potent inhibitor of GSK-3α and GSK-3β with IC50 values of 0.65 nM and 0.58 nM, respectively. | |||||
| Targets | GSK-3β | GSK-3α | p70 S6K | |||
| IC50 | 0.58 nM | 0.65 nM | >1 μM | |||
| Cell experiment: | |
Cell lines | Insulin receptor– expressing CHO-IR cells and primary rat hepatocytes |
Preparation method | The solubility of this compound in DMSO is <10 mm.="" general="" tips="" for="" obtaining="" a="" higher="" concentration:="" please="" warm="" the="" tube="" at="" 37="" °c="" for="" 10="" minutes="" and/or="" shake="" it="" in="" the="" ultrasonic="" bath="" for="" a="" while.stock="" solution="" can="" be="" stored="" below="" -20°c="" for="" several=""> |
Reaction Conditions | 24h; EC50=106 nM (CHO-IR cells); EC50=107 nM (rat hepatocytes). |
Applications | CHIR 98014 resulted in a stimulation of the GS activity ratio above basal. The concentrations of CHIR 98014 causing half-maximal GS stimulation (EC50) were 106 nM for CHO-IR cells and 107 nM for rat hepatocytes. |
| Animal experiment: | |
Animal models | Female db/db mice. |
Dosage form | 30 mg/kg; oral taken |
Applications | Markedly diabetic and insulin-resistant db/db mice treated with 30 mg/kg CHIR 98014 exhibited a significant reduction in fasting hyperglycemia within 4 h of treatment and showed improved glucose disposal during an IPGTT. |
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] Ring D B, Johnson K W, Henriksen E J, et al. Selective glycogen synthase kinase 3 inhibitors potentiate insulin activation of glucose transport and utilization in vitro and in vivo[J]. Diabetes, 2003, 52(3): 588-595. | |
CHIR-98014 Dilution Calculator
calculate
CHIR-98014 Molarity Calculator
calculate
| Cas No. | 252935-94-7 | SDF | Download SDF |
| Chemical Name | 6-N-[2-[[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-yl]amino]ethyl]-3-nitropyridine-2,6-diamine | ||
| Canonical SMILES | C1=CC(=C(C=C1Cl)Cl)C2=NC(=NC=C2N3C=CN=C3)NCCNC4=NC(=C(C=C4)[N+](=O)[O-])N | ||
| Formula | C20H17Cl2N9O2 | M.Wt | 486.31 |
| Solubility | ≥8.1mg/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. | ||
CHIR-98014 is a potent inhibitor of GSK-3α and GSK-3β with IC50 values of 0.65 nM and 0.58 nM, respectively [1]
GSK-3 (Glycogen synthase kinase 3) is a serine/threonine protein kinase and plays a pivotal role in a number of central intracellular signaling pathways, including cellular proliferation, migration, inflammation and immune responses, glucose regulation, and apoptosis. Recently, it has been reported that GSK-3 abnormally expressed in a variety of diseases, including Type II diabetes, Alzheimer"s Disease, inflammation, cancer, and bipolar disorder [2, 3].
CHIR-98014 is a potent GSK-3α and GSK-3β inhibitor. When tested with insulin receptor-expressing CHO-IR cells or primary rat hepatocytes, CHIR-98014 stimulated the GS activity ratio as high as two- to three fold compared with basal in a dose dependent manner. Similarly, in isolated type 1 skeletal muscle from insulin-sensitive lean Zucker and from insulin-resistant ZDF rats, administration of CHIR-98014 activated GS activity ratio [1]. In mouse ES-D3 cells, CHIR-98014 treatment (48 and 72 hours later) resulted in a significant activation of the Wnt/beta-catenin pathway via inhibiting GSK-3 [4].
In markedly diabetic and insulin-resistant db/db mice model, oral administration of CHIR-98014 (30mg/kg) significantly reduced fasting hyperglycemia within 4 hours and improved glucose disposal during an ipGTT [1].
References: [1].Ring, D.B., et al., Selective glycogen synthase kinase 3 inhibitors potentiate insulin activation of glucose transport and utilization in vitro and in vivo. Diabetes, 2003. 52(3): p. 588-95.[2]. Pan WA, et al. The RNA recognition motif of NIFK is required for rRNA maturation during cell cycle progression. RNA Biol. 2015. 12(3):255-67.[3]. McCubrey JA, et al. GSK-3 as potential target for therapeutic intervention in cancer. Oncotarget. 2014. 5(10):2881-911.[4]. Naujok O, et al. Cytotoxicity and activation of the Wnt/beta-catenin pathway in mouse embryonic stem cells treated with four GSK3 inhibitors. BMC Res Notes. 2014. 7(1):273-281.
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
细胞株(cell strain) 是通过选择法或克隆形成法从原代培养细胞中获得具有特殊性质或标志物的细胞称为细胞株。一般认为，细胞株是用单细胞分离培养或通过筛选的方法，由单细胞增殖形成的细胞群。细胞株的特殊性质或标志必须在整个培养期间始终存在。
细胞系(cell line) 是原代细胞经首次传代成功后即为细胞系。泛指一般可能传代的细胞。其中能够连续传代的细胞叫做连续细胞系或无限细胞系，不能连续培养的称为有限细胞系。大多数二倍体细胞为有限细胞系。由原先存在于原代培养物中的细胞世系所组成。如果不能继续传代，或传代次数有限， 可称为有限细胞系(finite cell line)， 如可以连续培养， 则称为连续细胞系(continuous cell line)， 培养50代以上并无限培养下去。人类肿瘤细胞，在体外培养半年以上，生长稳定，并连续传代的即可称为连续性株或系。
不知道楼主明白没有,意思是说培养基上的癌细胞可能过密,导致营养缺乏,无法满足生长分裂的需要,所以进入G0期,暂时不生长分裂.
3,MHC基因位于第六号染色体上,MHC存在于人类和哺乳动物的细胞表面.根据MHC所编码的分子结构和功能不同而将其分为1,2,3类?.1,2类分子结构相似,主要功能是向T细胞呈递抗原,是被T细胞识别的重要靶分子.1类存在于除红细胞以外的机体所以细胞膜上,2类分子仅存在于成熟B细胞,一些T细胞和抗原呈递细胞膜上,3类分子不具备上述功能,属于血清蛋白.
现在楼主应该明白了,MHC是不参与抗原抗体特异性结合的,是专门用来呈递抗原的.
哪种要好，有什么要求吗？接种多少代的细胞？
1新生大鼠鼠龄的选择新生大鼠心肌细胞在出生后3 d内具有部分的增殖能力，成年大鼠心肌细胞则为终末分化细胞，不再具有分裂增殖能力.因此，大鼠出生时间越短，其心肌细胞分离后成活率越高，越容易贴壁生长.大量观测表明，选择1~3 d龄大鼠分离其心肌细胞进行原代培养较为理想.其中尤以半日龄大鼠心肌细胞培养效果最佳.
2消化酶的选择及使用
新生大鼠心肌细胞的分离可采用组织块法和消化法，前者因不易获得密度均一的细胞且难控制成纤维细胞的生长而较少采用.消化法中常使用的酶有3种：胰蛋白酶、胶原酶I或Ⅱ以及透明质酸酶.透明质酸酶多与胰蛋白酶或胶原酶联合应用.胰蛋白酶作用较强，容易造成心肌细胞损坏.胶原酶作用较缓和，能消化细胞间质中的胶原纤维以释放细胞，对细胞损伤小，且在新生大鼠心肌组织，以胶原I为主,故我们选用胶原酶I.文献报道胶原酶的工作浓度一般在0.6~1 g・L1,我们使用的为0.8 g・L1.胶原酶最好现用现配.
3消化程度的把握
新生大鼠心肌细胞对酶消化极为敏感.消化过度可使肌原纤维出现萎缩，细胞死亡率增加或丧失贴壁能力及搏动能力;消化不足，细胞聚集成团，无法分清细胞边界，难以形态学观测.消化过程中使用磁力搅拌器时应注意1)转速一般控制在60~80 r・min1左右.(2)每次消化的时间须结合消化酶浓度确定.(3)将粘附在搅拌子上的心肌组织吹散，使酶液充分接触组织.(4)适宜温度为35~37℃.(5)当组织由红转白呈半透明状态时，应停止消化.
4接种的细胞密度
心肌细胞接种密度不仅影响细胞间的相互接触，进而影响细胞对肥大刺激的反应，而且影响长期培养细胞的成活率.接种细胞的绝对数量应经精确计算.一般而言，应根据实验的观测目的决定单位面积上的细胞数量.例如，如作形态学观测,六孔板中每孔的接种细胞数量应控制在1×105~2×105个;若需收获心肌细胞作mRNA或蛋白表达水平的观测，则每孔的接种密度可增加到5×105~6×105个.
原代细胞(primary cell) 是指从机体的组织(如人组织、小鼠组织、大鼠组织和兔组织等)经蛋白酶或其它的方法获得单个细胞并在体外进行模拟机体培养的细胞，称为原代细胞。一般认为，培养的原代的第1代细胞和传代到第10代以内的细胞统称为原代细胞培养。在人工条件下使其原代细胞生存、生长、繁殖和传代，进行细胞生命过程、细胞癌变、细胞工程等问题的研究。
细胞株(cell strain) 是通过选择法或克隆形成法从原代培养细胞中获得具有特殊性质或标志物的细胞称为细胞株。一般认为，细胞株是用单细胞分离培养或通过筛选的方法，由单细胞增殖形成的细胞群。细胞株的特殊性质或标志必须在整个培养期间始终存在。
细胞系(cell line) 是原代细胞经首次传代成功后即为细胞系。泛指一般可能传代的细胞。其中能够连续传代的细胞叫做连续细胞系或无限细胞系，不能连续培养的称为有限细胞系。大多数二倍体细胞为有限细胞系。由原先存在于原代培养物中的细胞世系所组成。如果不能继续传代，或传代次数有限， 可称为有限细胞系(finite cell line)， 如可以连续培养， 则称为连续细胞系(continuous cell line)， 培养50代以上并无限培养下去。人类肿瘤细胞，在体外培养半年以上，生长稳定，并连续传代的即可称为连续性株或系。
就好像撒一撮芝麻在培养基那么大的地方上，我们不能知道它会怎么落。
