| Cytochalasin Dinhibitor of actin polymerization, selective |
Sample solution is provided at 25 µL, 10mM.
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- Purity ≥95.00%
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Chemical structure
Related Biological Data
| Cell experiment [1-3]: | |
Cell lines | HeLa, Vero, L, HEp2, and MDBK cells, SC-1 cells, Murine CT26 colorectal carcinoma cells |
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 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months. |
Reacting condition | 0.2–0.5 μg/ml |
Applications | In HeLa, Vero, L, HEp2, and MDBK cells, cytochalasin D (0.2–0.5 μg/ml) induced sustained contraction (contracture), loss of microvilli, expression of endoplasmic contents (zeiosis), nuclear protrusion, and extension of cytoplasmic processes. Cells in G1 were most sensitive to CD; responsiveness decreased progressively during early S and is least in mid S through G2. CD inhibited transport of [14C]deoxyglucose in HeLa. In SC-1 cells, Cytochalasin D treatment severely disrupted network organization, increased the number of actin filament ends, and led to the formation of filamentous aggregates or foci composed mainly of actin filaments. Cytochalasin D (0.24~15 μg/mL, 16 h) inhibited CT26 tumor cell proliferation in time and dose dependent manner and induced significant CT26 cell apoptosis. |
| Animal experiment [3,4]: | |
Animal models | Murine CT26 tumor model, porcine coronary model |
Dosage form | Intravenous injection, 50 mg/kg, every 3 days for 21 days |
Application | Cytochalasin D (i.v., 50 mg/kg) in vivo treatment significantly inhibited tumor growth and prolonged the survival times in CT26 tumor-bearing mice. In porcine coronary model, Cytochalasin D (2 μg) resulted in less late lumen loss in low-dose. High-dose Cytochalasin D (20 μg) inhibited both late lumen loss and intimal area. |
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]. Miranda A F, Godman G C, Deitch A D, et al. Action of cytochalasin D on cells of established lines[J]. The Journal of cell biology, 1974, 61(2): 481-500. [2]. Schliwa M. Action of cytochalasin D on cytoskeletal networks[J]. The Journal of cell biology, 1982, 92(1): 79-91. [3]. Huang F Y, Li Y N, Mei W L, et al. Cytochalasin D, a tropical fungal metabolite, inhibits CT26 tumor growth and angiogenesis[J]. Asian Pacific journal of tropical medicine, 2012, 5(3): 169-174. [4].Salu K J, Bosmans J M, Huang Y, et al. Effects of cytochalasin D-eluting stents on intimal hyperplasia in a porcine coronary artery model[J]. Cardiovascular research, 2006, 69(2): 536-544. | |
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| Cas No. | 22144-77-0 | SDF | Download SDF |
| Synonyms | N/A | ||
| Chemical Name | (3S,3aR,4R,6R,6aS,7E,10R,12S,13Z,15R,15aS)-3-benzyl-6,12-dihydroxy-4,10,12-trimethyl-5-methylene-1,11-dioxo-2,3,3a,4,5,6,6a,9,10,11,12,15-dodecahydro-1H-cycloundeca[d]isoindol-15-yl acetate | ||
| Canonical SMILES | O[C@@H]1[C@@H](/C=C/C[C@@H](C)C2=O)[C@]3([C@@H](C=C[C@]2(C)O)OC(C)=O)[C@@H]([C@@H](C)C1=C)[C@H](CC4=CC=CC=C4)NC3=O | ||
| Formula | C30H37NO6 | M.Wt | 507.63 |
| Solubility | Soluble in DMSO | Storage | Desiccate at -20°C |
| Physical Appearance | A crystalline 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. | ||
Cytochalasin D is a selective inhibitor of actin polymerization with with IC50 value of 25 nM [1].Actin is a globular multi-functional protein and found nearly in all eukaryotic cells. it has been shown that actin polymerization plays a pivotal role in chemotaxis and cytokinesis. Cytochalasin D is reported as an inhibitor in the process of actin polymerization via disrupting actin microfilaments and activating p53-dependent pathways which in turn causes the arrest of cell cycle at the G1-S transition [2]. Cytochalasin D is a potent actin polymerization inhibitor. When tested with differentiating neurons, Cytochalasin D slowed down protrusion/retraction cycles and decreased lamellipodia axial motion via inhibiting actin polymerization [1]. In epithelial cell line HEp-2 cells, Cytochalasin D treatment regulated late and very late phases of viral transcription and shut down host transcription through blocking actin polymerization [3]. In the model of IPEC-J2 cells infected with PCV2, Cytochalasin D treatment could suppress PCV2 invasion, replication and release thus inhibited virus invasion [4].References: [1]. Sayyad, W.A., et al., The role of myosin-II in force generation of DRG filopodia and lamellipodia. Sci Rep, 2015. 5: p. 7842.[2]. Montazeri, M., et al., Activation of Toll-like receptor 3 reduces actin polymerization and adhesion molecule expression in endometrial cells, a potential mechanism for viral-induced implantation failure. Hum Reprod, 2015.[3]. Volkman, L.E., Baculoviruses and nucleosome management. Virology, 2015. 476c: p. 257-263.[4]. Yan, M., L. Zhu, and Q. Yang, Infection of porcine circovirus 2 (PCV2) in intestinal porcine epithelial cell line (IPEC-J2) and interaction between PCV2 and IPEC-J2 microfilaments. Virol J, 2014. 11: p. 193.
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培养一段时间后（一般小于一个细胞周期），固定细胞。根据所使用模拟核苷酸的不同，使用不同的方法。EdU，改变缓冲液的pH值，就会发出荧光。而BrdU则需用荧光抗体识别，需要对细胞膜和核膜进行通透处理。
再加入PI对DNA染色。
在二维坐标图上，横坐标设为线性PI荧光强度，纵坐标设为模拟核苷酸的荧光强度（log）。
典型的图就像下面一样：
G1, G2 和S期就按照图中的划分来做。G1期的细胞没有新合成DNA，所以BrdU信号是阴性，而PI的信号位于1倍体期。S期是正在合成DNA，所以BrdU都是阳性。G2期是已经合成好了2倍的DNA，所以位于2倍体期，信号是G1期的一倍。这个图中，G1 PI信号是300， G2就是600.
流式细胞仪（Flow cytometry ）是对细胞进行自动分析和分选的装置。它可以快速测量、存贮、显示悬浮在液体中的分散细胞的一系列重要的生物物理、生物化学方面的特征参量，并可以根据预选的参量范围把指定的细胞亚群从中分选出来。多数流式细胞计是一种零分辨率的仪器，它只能测量一个细胞的诸如总核酸量，总蛋白量等指标，而不能鉴别和测出某一特定部位的核酸或蛋白的多少。也就是说，它的细节 分辨率为零。
染色的各种染料，标记好的荧光抗体染色所需要的缓冲液，封闭液，PBS最终由于上样的缓冲液
不过还要考虑很多因素：标记的方法会不会影响细胞下面的培养细胞是否能够耐受长时间接触房间中的氧气和二氧化碳的浓度机器的无菌情况
（溶血）：红细胞，血红蛋白分解，红细胞溶解逃逸所述，称为溶血。通过各种物理和化学因素和毒素。在体外，如低渗溶液中，强烈的机械振荡，突然冷冻（-20℃-25℃）或突然化冻，过酸或过碱，以及乙醇，乙醚，皂碱，胆碱盐可引起溶血。人血浆的等渗溶液为0.9％NaCl溶液，红细胞在低于0.45％的NaCl溶液中，由于水的渗透，肿胀和红细胞破裂，血红蛋白逸出。在体内，溶血溶血性细菌侵入或某些毒素的抗原 - 抗体反应（如输入配血不合的血液），各种机械性损伤，红细胞内在（膜，酶）引起某些药物的缺陷。溶血性细菌，如某些溶血性链球菌和产气荚膜梭菌可导致败血症。的红血细胞和某些溶血性毒液含酶卵磷脂，卵磷脂血浆或红细胞成溶血卵磷脂，使红细胞膜分解疟原虫破坏。
比较常用的是用计数微球。比如临床上的CD4细胞绝对计数，就是采用这个方法。试管内事先已经有固定数目的计数微球和染色抗体。按要求加入固定体积的抗凝血液，裂解红细胞后上机。记录一千个微球，同时就会得到不同染色区域细胞的数量。这样就可以算出单位体积血液内CD4细胞的绝对数目了。
这样的计数微球也可以另外购买。浓度是固定的。在你已知体积的细胞悬液中加入一定体积的微球后上机，记录一千个微球的数据，细胞的计数也会同时记录。根据微球的浓度就可以推算出细胞的浓度，从而得出绝对计数了
另外一个方法是有些流式细胞仪是使用微泵加样，而不是连续吸取样本，这样样本的体积是已知的，记录一次就是所有样本的数量，可以算出细胞浓度
