(-)-Blebbistatin

For research use only. Not for use in humans.

目录号:S7099 别名: (S)-(-)-Blebbistatin 中文名称:布雷他汀

(-)-Blebbistatin Chemical Structure

CAS No. 856925-71-8

(-)-Blebbistatin ((S)-(-)-Blebbistatin)是一种细胞渗透性抑制剂,作用于非肌球蛋白IIATPase,无细胞试验中IC50为~2 μM,不抑制肌球蛋白轻链激酶 (MLCK),抑制卵裂沟的缢缩,不干扰有丝分裂或收缩环的组装。

规格 价格 库存 购买数量  
10mM (1mL in DMSO) RMB 1792.87 现货
RMB 1385.26 现货
RMB 3023.26 现货
RMB 7444.71 现货
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客户使用Selleck生产的(-)-Blebbistatin发表文献32篇:

产品安全说明书

ATPase抑制剂选择性比较

生物活性

产品描述 (-)-Blebbistatin ((S)-(-)-Blebbistatin)是一种细胞渗透性抑制剂,作用于非肌球蛋白IIATPase,无细胞试验中IC50为~2 μM,不抑制肌球蛋白轻链激酶 (MLCK),抑制卵裂沟的缢缩,不干扰有丝分裂或收缩环的组装。
靶点
non-muscle myosin II ATPases [2]
(Cell-free assay)
0.5 μM-5 μM
体外研究

Blebbistatin抑制细胞分裂,改变鱼角膜细胞的平滑运动,且抑制缺乏细丝蛋白的细胞系进行自发起泡。Blebbistatin抑制非肌肉肌球蛋白IIA,非肌肉肌球蛋白IIB,和兔骨骼肌肌球蛋白S1的HMM片段的酶活性,而不抑制平滑肌球蛋白。[1]Blebbistatin快速且可逆抑制Mg-ATPase活性,也抑制非肌肉肌球蛋白IIA和IIB的体外活性,而极其微弱抑制平滑肌肌球蛋白(IC50=80 μM)。Blebbistatin有效抑制Dictyostelium肌球蛋白II,但极其微弱抑制Acanthamoeba肌球蛋白II。Blebbistatin不抑制代表性的I,V,和X型肌球蛋白超家族成员。[2]Blebbistatin不与核苷酸竞争性结合到骨骼肌肌球蛋白亚片段-1。Blebbistatin优先结合到ATP酶,与ADP和磷酸盐在活性位点相互作用,减慢磷释放。Blebbistatin既不干扰肌球蛋白与肌动蛋白结合,也不干扰ATP诱导的肌动球蛋白解离。[3]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Rh41 NGLQ[XZyUFSVIHHzd4F6 MVTxTHRUKG:oIIDl[IlifHKrYzDjZY5k\XJiY3XscEBtcW6nczD0c{Bq\GWwdHnmfUBufWy2aYDs[UBweHCxcoT1col1cWW|IH\vdkBlenWpIILldJVzeG:|aX7nPkBEd26oaYLtZZRwenlic3Py[YVvKG[xcjDSbFQyKGOnbHzz M2T0SFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ7NEO1NVM6Lz5{OUSzOVE{QTxxYU6=
SK-N-MC NXzaNnY2eUiWUzDhd5NigQ>? M4LmWJFJXFNib3[gdIVlcWG2cnnjJINidmOncjDj[YxtKGyrbnXzJJRwKGmmZX70bYZ6KG23bITpdIxmKG:ycH;yeJVvcXSrZYOg[o9zKGS{dXegdoVxfXKyb4Ppcoc7KFC{aX3hdpkhe2O{ZXXuJIZweiCVSz3OMW1EKGOnbHzz NV\XTFVGRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkm0N|UyOzlpPkK5OFM2OTN7PD;hQi=>
NB-EBc1 NXPEbYtveUiWUzDhd5NigQ>? NHmyem9yUFSVIH;mJJBm\GmjdILpZ{Bk[W6lZYKgZ4VtdCCuaX7ld{B1dyCrZHXueIlngSCvdXz0bZBt\SCxcIDvdpR2dmm2aXXzJIZweiCmcoXnJJJmeHW{cH;zbY5oQiCScnntZZJ6KHOlcnXlckBnd3JiTlKtSWJkOSClZXzsdy=> MYm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zQTR|NUGzPUc,Ojl2M{WxN|k9N2F-

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
talin 1 / vinculin / paxillin; 

PubMed: 20308429     


Effects of myosin II inhibition on the interactions between vinculin, paxillin, and talin. IPs were performed from lysates of untreated control MEFs (control) or MEFs treated with 20 µM blebbistatin (Blebb) followed by immunoblot analysis. (A) IP with anti-vinculin (Vcl) antibodies, immunoblot with anti-vinculin and antipaxillin (Pxn; left), or anti–talin 1 (Tln; right) antibodies. (B) IP with antipaxillin antibodies and immunoblotting with antivinculin antibodies. White lines indicate that intervening lanes have been spliced out. (C) IP with anti–talin 1 antibodies and immunoblotting with antivinculin antibodies are shown.

paxillin / pY31 paxillin / pY397FAK / FAK; 

PubMed: 20308429     


(A) Immunoblot analysis of lysates of untreated (control) and cells treated with 20 µM blebbistatin (Blebb) or with 20 µM blebbistatin and 100 µM Na3VO4 using antibodies specific to paxillin (Pxn), pY31 paxillin, pY397FAK, or FAK.

PY epitopes / vinculin / paxillin; 

PubMed: 20308429     


(D) Antipaxillin IPs from lysates of untreated control or cells treated with either 20 µM blebbistatin, 100 µM Na3VO4, and 20 µM blebbistatin or 100 µM Na3VO4 alone followed by analysis by PAGE and immunoblotting with antibodies to vinculin, paxillin, or PY epitopes. (E–I) EGFP-conjugated paxillin (Pxn-GFP wt) or paxillin bearing mutations of tyrosines 31 and 118 to phenylalanines (Pxn Y31/118F) or glutamic acids (Pxn Y31/118E) were expressed in MEFs and either treated with 20 µM blebbistatin or not.

20308429
Growth inhibition assay
Cell viability; 

PubMed: 26733241     


(B) The time course of cell viability of isolated ventricular mouse myocytes in culture at 2, 24, and 48 h postplating (representative images). Supplementation with blebbistatin alone maintains cell viability to a greater extent that culture with BDM, blebbistatin and BDM in combination, or without any supplementation at both 24 and 48 h.

Cell death; 

PubMed: 26733241     


(C) Viability of myocytes assessed by PI staining at 6, 18, 24, and 48 h after plating. Supplementation with blebbistatin keeps a greater proportion of isolated mouse myocytes alive throughout the 48 h post isolation. *versus control P < 0.05.

26733241
Glycerol/urea gel electrophoresis
RLC phosphorylation; 

PubMed: 18701651     


A: cultures were incubated with 100 nM KT5926 (KT) for 60 min, and RLC phosphorylation was analyzed by glycerol/urea gel electrophoresis. Inhibition of MLCK caused a significant reduction in constitutive RLC phosphorylation. Monolayers preincubated with 100 nM KT5926 for 30 min and then treated with blebbistatin for an additional 30 min in the presence of inhibitor showed that MLCK was responsible for blebbistatin-induced RLC phosphorylation.

RLC phosphorylation; 

PubMed: 18701651     


B: effect of blebbistatin on MLCK catalytic activity. Both MLCK210 (lane 1) and MLCK155 (lane 4) catalyze RLC phosphorylation in the presence of Ca2+/CaM, whereas in the absence of Ca2+/CaM, no 32PO4 was incorporated into myosin II RLCs (lanes 2 and 5). Blebbistatin does not directly activate MLCK since incubation of either isoform of MLCK in phosphorylation buffer containing 50 μM blebbistatin in the absence of Ca2+/CaM resulted in no RLC phosphorylation (lanes 3 and 6). Blebbistatin does not inhibit MLCK activity in the presence of Ca2+/CaM (lanes 7 and 8). Lane 1, MLCK210 + Ca2+/CaM; lane 2, MLCK210 + EGTA; lane 3, MLCK210 + EGTA + 50 μM blebbistatin; lane 4, MLCK155 + Ca2+/CaM; lane 5, MLCK155 + EGTA; lane 6, MLCK155 + EGTA + 50 μM blebbistatin; lane 7, MLCK210 + Ca2+/CaM + 50 μM blebbistatin; lane 8, MLCK155 + Ca2+/CaM + 50 μM blebbistatin.

RLC phosphorylation; 

PubMed: 18701651     


C: effect of Ca2+ depletion on blebbistatin-induced RLC phosphorylation. For Ca2+ chelation, monolayers were preincubated for 1 h at room temperature in Ca2+-free HBSS containing 10 mM BAPTA-AM/1 μM thapsigargin. Cells were treated with blebbistatin for 30 min; samples were separated by glycerol/urea gel electrophoresis, transferred to nitrocellulose, and probed with anti-myosin RLC antibody. Ca2+ depletion prevents blebbistatin-induced RLC phosphorylation.

18701651
DIC image
Traction force of a palladin KD (Palld4) cell; 

PubMed: 27353427     


Palladin knockdown cells show more efficient recovery from Blebbistatin treatment. (A) DIC image of a palladin KD (Palld4) cell on a gel of intermediate stiffness (10–30 kPa range). Scale bar: 10 μm. (B) DIC image of the same cell as in A, 30 minutes after incubation in 15 μM blebbistatin. (C) DIC image of the cell 1 hour after washout from blebbistatin, showing recovery of cell morphology. (D) Traction force map of the cell in A showing robust generation of traction forces. (E) Traction force map of the cell in B, showing disappearance of traction forces upon blebbistatin addition. (F) Traction force map of the cell in C, showing recovery of traction forces 1 hour after Blebbistatin washout.

27353427
Immunofluorescence
GCs morphology; 

PubMed: 25598228     


The effect of Blebbistatin on GCs morphology. (a–b) Lamellipodium emerging from a DRG neuron in control condition and after treatment with 30 μM Blebbistatin, respectively. Note the 'filopodish' appearance of the lamellipodia after Blebbistatin treatment. (c) Immunostaining of DRG lamellipodium in control condition for actin (green) and tubulin (blue) staining. (d) As in (c) but in the presence of 30 μM Blebbistatin.

actin / NMIIA / tubulin; 

PubMed: 25598228     


(e) Immunostaining of a GC after Blebbistatin treatment for actin, NMIIA and tubulin and merge of the three staining. Arrows and arrowheads indicate filopodia with and without a clear staining for tubulin, respectively. (f) Immunostaining of a GC after Blebbistatin treatment for actin, NMIIB and tubulin and merge of the three staining.

VE-cadherin / F-actin; 

PubMed: 22090347     


(B) Immunofluorescence of VE-cadherin (red) and phalloidin staining of F-actin (green) following 30-min treatment with control inactive (+)-blebbistatin (left) or active (−)-blebbistatin (right) reveal reduced size of VE-cadherin puncta following myosin II inhibition. Insets are magnified with equal ratios relative to boxed regions in original image.

PY epitopes / actin; 

PubMed: 20308429     


Rho kinase–mediated myosin II activity and substrate stiffness slow MEF migration and increase adhesion size.(C) Immunolocalization of PY epitopes (P-Tyr) to visualize adhesions (green) and fluorescent phalloidin staining to visualize actin filaments (red) under the treatments as untreated cells (control), cells treated with 20 µM blebbistatin (Blebb) or 10 µM Y27632, or plated on 1.0 kPa compliant polyacrylamide substrates. Merged images are shown in the third column, and boxed regions are magnified in the fourth column. Bars: (third column) 10 µm; (fourth column) 2 µm.

PY epitopes / paxillin; 

PubMed: 20308429     


(A) Immunolocalization of paxillin (Pxn; red) and PY epitopes (P-Tyr; green) in untreated cells (control) or cells treated with 20 µM blebbistatin (Blebb). Merged images are shown in the third column, and boxed regions are magnified in the fourth column. Bars: (third column) 10 µm; (fourth column) 2 µm.

talin 1 / FAK / β1 integrin / zyxin / vinculin / α-actinin / paxillin; 

PubMed: 20308429     


(B) Immunolocalization of paxillin (red) with either (in green); talin 1 (Tln), FAK, β1 integrin (β1-Int), zyxin (Zyx), vinculin (Vcl), or α-actinin (Actn) in untreated control cells (left) or cells treated with 20 µM blebbistatin (right). Merged images are shown in the right column. Bar, 2 µm.

25598228 22090347 20308429

推荐的实验操作(此推荐来自于公开的文献所以Selleck并不保证其有效性)

溶解度 (25°C)

体外 DMSO 58 mg/mL (198.4 mM)
Water Insoluble
Ethanol Insoluble
体内 从左到右依次将纯溶剂加入产品,现配现用(数据来自Selleck实验检测而非文献):
2% DMSO+40% PEG 300+5% Tween 80+ddH2O
5mg/mL

* 溶解度检测是由Selleck技术部门检测的,可能会和文献中提供的溶解度有所差异,这是由于生产工艺和批次不同产生的正常现象。请按照顺序依次加入各个纯溶剂。

化学数据

分子量 292.33
化学式

C18H16N2O2

CAS号 856925-71-8
储存条件 3年 -20°C(避光) 粉状
2年 -80°C(避光) 溶于溶剂
别名 (S)-(-)-Blebbistatin

动物体内配方计算器 (澄清溶液)

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
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Selleck为您提供正确的澄清溶液配方)
% DMSO % % Tween 80 % ddH2O
计算重置

计算器

摩尔浓度计算器

摩尔浓度计算器

本计算器可帮助您计算出特定溶液中溶质的质量、溶液浓度和体积之间的关系,公式为:

质量 (mg) = 浓度 (mM) x 体积 (mL) x 分子量 (g/mol)

摩尔浓度计算公式

  • 质量
    浓度
    体积
    分子量

*在配置溶液时,请务必参考Selleck产品标签上、SDS / COA(可在Selleck的产品页面获得)批次特异的分子量使用本工具。

稀释计算器

稀释计算器

用本工具协助配置特定浓度的溶液,使用的计算公式为:

开始浓度 x 开始体积 = 最终浓度 x 最终体积

稀释公式

稀释公式一般简略地表示为: C1V1 = C2V2 ( 输入 输出 )

  • C1
    V1
    C2
    V2

在配置溶液时,请务必参考Selleck产品标签上、SDS / COA(可在Selleck的产品页面获得)批次特异的分子量使用本工具。.

连续稀释计算器方程

  • 连续稀释

  • 计算结果

  • C1=C0/X C1: LOG(C1):
    C2=C1/X C2: LOG(C2):
    C3=C2/X C3: LOG(C3):
    C4=C3/X C4: LOG(C4):
    C5=C4/X C5: LOG(C5):
    C6=C5/X C6: LOG(C6):
    C7=C6/X C7: LOG(C7):
    C8=C7/X C8: LOG(C8):
分子量计算器

分子量计算器

通过输入化合物的化学式来计算其分子量:

总分子量:g/mol

注:化学分子式大小写敏感。C10H16N2O2 c10h16n2o2

摩尔浓度计算器

质量 浓度 体积 分子量
计算

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID