Powder: -20°C for 3 years | In solvent: -80°C for 2 years
Thiazovivin 是一种ROCK 抑制剂,能提高 iPSC 的生成效率,对人胚胎干细胞具有保护作用。
产品描述 | Thiazovivin, a ROCK inhibitor (IC50: 0.5 μM), increases the survival rate of hESC. |
靶点活性 | ROCK:~0.5 μM (cell free) |
体外活性 | When the cells were cultured in the presence of an E-cadherin blocking antibody, the formation of large, compact aggregates following Thiazovivin treatment was severely inhibited and extensive cell death was observed. Thiazovivin (2 μM) inhibits ROCK activity and protects hESCs at a similar level as Y-27632 (10 μM), a widely used selective ROCK inhibitor [1]. Attachment rates of blastocyst and embryonic cell clumps onto feeder cells in the Thiazovivin treatment group were greater than those of the control group. The pluripotency markers of the OCT4 and NANOG genes and the adhesion molecule E-cadherin were increased by Thiazovivin treatment [2]. By adding thiazovivin to reprogramming cultures, the reprogramming efficiency of CB cells increases by more than 10 times [3]. |
激酶实验 | Synthesis of compound immobilized affinity matrixes: Compounds Tzv and its inactive analog (10 mg each) in DMSO (500 μL) and Et3N (10.4 μL) were added to Reacti-Gel (0.5 mL, 25 μmol) that was washed by DMSO in an Eppendorf vial. The reaction mixture was incubated at room temperature until the starting material disappeared (determined by HPLC). After the disappearance of the starting material, ethanolamine (15 μL) was added and the resulting mixture was incubated at room temperature overnight to block the Reacti-Gel. The resulting affinity matrices were washed thoroughly with DMSO (500 μL × 4), PBS (500 μL × 2), and stored at 4 °C in NaN3 solution (0.1% in PBS). Affinity pull-down was performed as described previously. Briefly, whole cell lysates were pretreated with the unfunctionalized affinity matrix at 4 °C for 1.5 h. After washing three times, samples were incubated with the positive or negative affinity matrix at 4 °C for 1 h. After heat shock, samples were loaded and separated on a 4–20% Tris-Glycine SDS PAGE and silver stained with a Silver Stain Plus Kit. The differentially retained protein bands were cut, destained, and analyzed with LCMS [1]. |
细胞实验 | Chemically defined and feeder-free human embryonic stem cell (hESC) culture was described briefly as following. hESCs were grown on Matrigel-coated tissue culture plates in N2B27-CDM [DMEM-F12 supplemented with 1× N2 supplements, 1× B27 supplements, 2 mM L glutamine, 0.11 mM 2-mercaptoethanol, 1× nonessential amino acids, and 0.5 mg/mL BSA (fraction V)] and 20 ng/mL bFGF. Human ESCs were passaged every five to six days with 0.05% trypsin. Murine ESCs are cultured in knockout DMEM supplement with 2 mM L glutamine, 1× nonessential amino acids, 15% serum replacement, and 1 × 10^3 ng/mL leukemia inhibitory factor (LIF). For clonal survival assays, single hESCs were diluted to clonal density and plated onto 96-well Matrigel-coated plate. For low-density survival assays, 500 cells were plated onto 96-well Matrigel-coated plate. To visualize hESC colonies, cultures were fixed in 4% paraformaldehyde in phosphate buffered saline (PBS) for 5 min, washed once in PBS, then stained for alkaline phosphatase (ALP) activity as described in the manufacturer's instructions. ALP-positive colonies were counted on an inverted microscope. For growing hESCs in mouse medium, HES2, HUES7, HUES9, and HUES1-Oct4-GFP were cultured in murine ESC (mESC) growth media supplemented with 1-μM mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) inhibitor PD0325901 and 5-μM p38 inhibitor SB202190 and 1 × 10^3 human LIF [1]. |
分子量 | 311.36 |
分子式 | C15H13N5OS |
CAS No. | 1226056-71-8 |
Powder: -20°C for 3 years | In solvent: -80°C for 2 years
DMSO: 31.1 mg/mL (100 mM)
( < 1 mg/mL refers to the product slightly soluble or insoluble )
对于不同动物的给药剂量换算,您也可以参考 更多...
请在以下方框中输入您的动物实验信息后点击计算,可以得到母液配置方法和体内配方的制备方法: 比如您的给药剂量是10 mg/kg,每只动物体重20 g,给药体积100 μL,一共给药动物10 只,您使用的配方为5% DMSO+30% PEG300+5% Tween 80+60% ddH2O。那么您的工作液浓度为2 mg/mL。
母液配置方法:2 mg 药物溶于 50 μL DMSO (母液浓度为 40 mg/mL), 如您需要配置的浓度超过该产品的溶解度,请先与我们联系。
体内配方的制备方法:取 50 μL DMSO 主液,加入 300 μL PEG300, 混匀澄清,再加 50 μL Tween 80,混匀澄清,再加 600 μL ddH2O, 混匀澄清。
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您可能有的问题的答案可以在抑制剂处理说明中找到,包括如何准备库存溶液,如何存储产品,以及基于细胞的分析和动物实验需要特别注意的问题。
Thiazovivin 1226056-71-8 Cell Cycle/Checkpoint Cytoskeletal Signaling Stem Cells ROCK Inhibitor Rho-associated protein kinase ROK Rho-associated kinase inhibit Rho-kinase inhibitor