CTX0294885
目录号 : GC17637
A broad spectrum kinase inhibitor used in proteomics
Cas No.:1439934-41-4
Sample solution is provided at 25 µL, 10mM.
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CTX-0294885 is a a novel bisanilino pyrimidine; exhibits inhibitory activity against a broad range of kinases in vitro, and further developed it into a Sepharose-supported kinase capture reagent.Target: Kinase capture reagentUse of a quantitative proteomics approach confirmed the selectivity of CTx-0294885-bound beads for kinase enrichment. Large-scale CTx-0294885-based affinity purification followed by LC-MS/MS led to the identification of 235 protein kinases from MDA-MB-231 cells, including all members of the AKT family that had not been previously detected by other broad-spectrum kinase inhibitors. Addition of CTx-0294885 to a mixture of three kinase inhibitors commonly used for kinase-enrichment increased the number of kinase identifications to 261, representing the largest kinome coverage from a single cell line reported to date. Coupling phosphopeptide enrichment with affinity purification using the four inhibitors enabled the identification of 799 high-confidence phosphosites on 183 kinases, ～10% of which were localized to the activation loop, and included previously unreported phosphosites on BMP2K, MELK, HIPK2, and PRKDC. Therefore, CTx-0294885 represents a powerful new reagent for analysis of kinome signaling networks that may facilitate development of targeted therapeutic strategies.
References:
[1]. Zhang L, et al. Characterization of the novel broad-spectrum kinase inhibitor CTx-0294885 as an affinity reagent for mass spectrometry-based kinome profiling. J Proteome Res. 2013 Jul 5;12(7):3104-16.
| Kinase experiment [1]: | |
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Inhibitory assays |
A panel of 10 kinases consisting of FAK, Flt3, JAK2, JAK3, CSF-1R, SRC, VEGFR3, Aurora A, LIMK1, and ROCK2 was initially used to evaluate kinase inhibition profiles. 15 μL assay reactions were performed in Greiner brand white 384-well, low-volume plates. All reactions contained assay buffer (10 mM HEPES pH 7.4, 10 mM MgCl2, 0.01% (v/v) Tween-20, 50 μM Na3VO4, 0.01% (w/v) albumin from chicken egg white, 1 mM dithiothreitol), a N-terminally biotinylated peptide substrate (concentration 111 nM), and ATP at a concentration equaling the KM (ATP) of the kinase. The compounds were added in a volume of 100 nL from an 11-point-dilution series prepared in DMSO, positive and negative control reactions receiving the same volume of DMSO without compound. The kinases were added at predetermined concentrations, generally ranging between 0.2 and 8 nM, with the enzyme being omitted from negative control reactions. The reactions were incubated for 90 min at 30 °C and stopped by adding 5 μL of Stop buffer (10 mM HEPES pH 7.4, 25 mM NaCl, 100 mM EDTA, 0.01% (v/v) Tween-20) containing streptavidin-coated donor and antiphosphotyrosine (P-Tyr-100) acceptor beads. Plates were incubated for 4–6 h before being read on a PerkinElmer EnVision plate reader in HTS Alphascreen mode. |
| Cell experiment [1]: | |
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Cell lines |
MDA-MB-231 Cells |
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Preparation method |
The solubility of this compound in DMSO is > 21.9 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. |
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Reacting condition |
1 μM |
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Applications |
CTX-0294885 exhibited inhibitory activity against a broad range of kinases in vitro, and further developed it into a Sepharose-supported kinase capture reagent. Large-scale CTx-0294885-based affinity purification followed by LC-MS/MS led to the identification of 235 protein kinases from MDA-MB-231 cells, including all members of the AKT family that had not been previously detected by other broad-spectrum kinase inhibitors. |
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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. |
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References: [1]. Zhang L, Holmes I P, Hochgrafe F, et al. Characterization of the novel broad-spectrum kinase inhibitor CTx-0294885 as an affinity reagent for mass spectrometry-based kinome profiling[J]. Journal of proteome research, 2013, 12(7): 3104-3116. | |
| Cas No. | 1439934-41-4 | SDF | |
| 化学名 | (Z)-2-((E)-(5-chloro-2-((4-(piperazin-1-yl)phenyl)amino)pyrimidin-4(3H)-ylidene)amino)-N-methylbenzimidic acid | ||
| Canonical SMILES | C/N=C(O)/C1=CC=CC=C1/N=C(N2)\C(Cl)=CN=C2NC3=CC=C(N4CCNCC4)C=C3 | ||
| 分子式 | C22H24ClN7O | 分子量 | 437.93 |
| 溶解度 | ≥ 21.9mg/mL in DMSO | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.2835 mL | 11.4173 mL | 22.8347 mL |
| 5 mM | 0.4567 mL | 2.2835 mL | 4.5669 mL |
| 10 mM | 0.2283 mL | 1.1417 mL | 2.2835 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet