1-NM-PP1
(Synonyms: 4-氨基1-叔丁基-3-(1'-萘甲基)吡唑并[3,4-D]嘧啶,PP1 Analog II) 目录号 : GC11696
Inhibitor of modified 'analog-sensitive' kinases
Cas No.:221244-14-0
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >99.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Kinase experiment [1]: | |
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In vitro kinase assays |
We carried out in vitro kinase assays (except for Cdc28) in the presence of 0.2 μCi /μl [γ-32P]ATP at low ATP concentration (10 nM) so that IC50 values represent a rough measure of the inhibition constant (Ki). Measurement of inhibitor IC50 values were done as Liu, Y. et al. described. Purified Cdc28–His6 (1 nM) and MBP–Clb2 (3 nM) were incubated for 10 min at 23℃ in a 25 μl reaction mixture containing 5 μg histone H1, 1 μCi of [γ-32P]ATP (1 μCi per 10 μM and 1μCi per 1 mM), and varying concentrations of 1-NM-PP1 in kinase buffer (25mM HEPES-NaOH pH 7.4, 10mM NaCl, 10mM MgCl2 and 1mM dithiothreitol). Reaction products were analysed by 15% SDS–PAGE followed by autoradiography. For the determination of Cdc28 kinetic constants, varying concentrations of [γ-32P]ATP (1 μCi per 100 μM) were incubated and analysed as above. |
| Cell experiment [2]: | |
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Cell lines |
PDK1?/? ES cells |
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Preparation method |
This compound is soluble in DMSO. 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 |
10 μM for 24h-72h; or 20 μM for 48h |
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Applications |
1-NM-PP1 suppressed the phosphorylation of PDK1 targets in PDK1?/? ES cells expressing PDK1 L159G but not WT PDK1. Moreover, 1-NM-PP1 inhibited sorbitol-induced MSK1 S212 T-loop phosphorylation, ERK/p38MAPK phosphorylation sites S581 and ERK/p38 dependent autophosphorylation at S376. 1-NM-PP1 also inhibited IGF1-stimulated PKB/Akt T308 phosphorylation and S6K activity in PDK1?/? +LG ES cells. |
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References: 1. Bishop, A. C., Ubersax, J. A., Petsch, D. T., Matheos, D. P., Gray, N. S., Blethrow, J., Shimizu, E., Tsien, J. Z., Schultz, P. G., Rose, M. D., Wood, J. L., Morgan, D. O. and Shokat, K. M. (2000) A chemical switch for inhibitor-sensitive alleles of any protein kinase. Nature. 407, 395-401 2. Tamguney, T., Zhang, C., Fiedler, D., Shokat, K. and Stokoe, D. (2008) Analysis of 3-phosphoinositide-dependent kinase-1 signaling and function in ES cells. Exp Cell Res. 314, 2299-2312 | |
1-NM-PP1 is a C3-modified analog of cell-permeable PP1, acts as a potent and selective inhibitor of protein kinase families [1].
1-NM-PP1 has shown the selective inhibition of protein kinases that have been mutated with the IC50 values of 28 μM, 1.0 μM, 3.4 μM, 29 μM, 24 μM,4.3 nM, 3.2 nM, 120 nM,5 nM and 8 nM for v-Src, c-Fyn, c-Abl, CDK2, CAMKII, v-Src-as1, c-Fyn-as1, cAbl-as2, CDK2-as1 and CAMKII-as1, respectively. 1-NM-PP1 inhibited each of the five target kinases at low nanomolar concentrations with target-specificities ranging from 85-fold to 400-fold measured against the most inhibitable wild-type kinases. In addition, the in vitro study has also revealed that Cdc28-as1 is highly sensitive to 1-NM-PP1 with ATP kinetic Km values of 35 μM and 322 μM, the Kcat values of 132 min-1 and 21.3 min-1, the Kcat/Km values of 3.7 and 0.066, the IC50 values of 22 μM and 0.002 μM for Cdc28/Clb2 and Cdc28-as1/Clb2, respectively [1].
References:
[1] Bishop AC1, Ubersax JA, Petsch DT, Matheos DP, Gray NS, Blethrow J, Shimizu E, Tsien JZ, Schultz PG, Rose MD, Wood JL, Morgan DO, Shokat KM. A chemical switch for inhibitor-sensitive alleles of any protein kinase. Nature. 2000 Sep 21; 407(6802):395-401.
| Cas No. | 221244-14-0 | SDF | |
| 别名 | 4-氨基1-叔丁基-3-(1'-萘甲基)吡唑并[3,4-D]嘧啶,PP1 Analog II | ||
| 化学名 | 1-tert-butyl-3-(naphthalen-1-ylmethyl)pyrazolo[3,4-d]pyrimidin-4-amine | ||
| Canonical SMILES | CC(C)(C)N1C2=C(C(=N1)CC3=CC=CC4=CC=CC=C43)C(=NC=N2)N | ||
| 分子式 | C20H21N5 | 分子量 | 331.41 |
| 溶解度 | ≥ 16.55mg/mL in DMSO, ≥ 9.22 mg/mL in EtOH with ultrasonic and warming | 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 | 3.0174 mL | 15.0871 mL | 30.1741 mL |
| 5 mM | 0.6035 mL | 3.0174 mL | 6.0348 mL |
| 10 mM | 0.3017 mL | 1.5087 mL | 3.0174 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。