肌動蛋白是一類形成微絲的球狀多功能蛋白質(zhì) 。它基本上存在于所有真核細(xì)胞中 ，進(jìn)化過程中高度保守。細(xì)胞肌動蛋白以單體和聚合體形式存在于真核細(xì)胞內(nèi)，其中單體小球又稱為G-肌動蛋白（Globular Actin，G-actin）；多個(gè)G-肌動蛋白以雙螺旋結(jié)構(gòu)聚合形成纖維化肌動蛋白，稱為F-肌動蛋白（Fibros Actin，F(xiàn)-actin），又稱為微絲，是細(xì)胞骨架最主要的結(jié)構(gòu)組分之一。

G-Actin/F-Actin這2種形式在一定生理?xiàng)l件下相互轉(zhuǎn)換，在微絲正極組裝和負(fù)極去組裝同時(shí)存在的現(xiàn)象，此現(xiàn)象被命名為“踏車現(xiàn)象”（TreadMilling），并在細(xì)胞內(nèi)保持平衡狀態(tài)，參與細(xì)胞一系列的生理功能。當(dāng)G-Actin/F-Actin在細(xì)胞內(nèi)比值發(fā)生改變時(shí)，意味著細(xì)胞結(jié)構(gòu)，運(yùn)動形態(tài)和穩(wěn)態(tài)的變化。

因此G-Actin/F-Actin比值的檢測，常被用于：

1.研究藥物化合物對細(xì)胞穩(wěn)態(tài)的影響。

2.研究突變的細(xì)胞系與野生型細(xì)胞的變化。

3.研究環(huán)境的變化對細(xì)胞的應(yīng)激。

作為專業(yè)的生命科學(xué)醫(yī)藥原料供應(yīng)商，CytoSkeleton中國區(qū)金牌代理，艾美捷科技為您推薦：G-Actin/F-Actin比值分析試劑盒( G-Actin：F-Actin 細(xì)胞體內(nèi)分析試劑盒 )

* 本產(chǎn)品僅適用于科研用途.

常見問題FAQ：

1. G-Actin/F-Actin比值分析試劑盒的實(shí)驗(yàn)中，進(jìn)行到哪一步，可暫停實(shí)驗(yàn)？

直到在37°C下以100,000xg旋轉(zhuǎn)1小時(shí)后才能停止測定。高速離心后，可以將上清液（G-肌動蛋白）與SDS上樣緩沖液混合并冷凍以備后用。沉淀物（F-肌動蛋白）應(yīng)該用解聚劑和水重新懸浮，然后與SDS上樣緩沖液混合并冷凍以備后用。冷凍后，F(xiàn)-肌動蛋白解聚，因此有必要在冷凍樣品之前將F-肌動蛋白與G-肌動蛋白分離，以分離樣品以準(zhǔn)確測量F-肌動蛋白和G-肌動蛋白的比率。

2. G-Actin/F-Actin比值分析試劑盒的靈敏度多高？

該測定可以檢測到G-肌動蛋白與F-肌動蛋白比率的小至15%的變化。每個(gè)條件都應(yīng)一式兩份進(jìn)行并重復(fù)多次，因?yàn)閷?shí)驗(yàn)之間的檢測重現(xiàn)性可能會有10-20%的差異。

3. G-Actin/F-Actin比值分析試劑盒能否在較慢的離心速度（如16,000g）下工作？

抱歉不行。在我們的測試離心速度是發(fā)現(xiàn)，如果低于100,000xg，哪怕是26,000 g，未能有效地沉淀F-肌動蛋白。

結(jié)果展示：

*以上結(jié)果僅供參考，因細(xì)胞系的不同，細(xì)胞所處的運(yùn)動狀態(tài)，藥物與環(huán)境刺激不同，G-actin/F-actin的比值會有較大的浮動，建議參考相同細(xì)胞系的已發(fā)表文獻(xiàn)。

部分發(fā)表文章展示：

• Morandell, Jasmin, et al. "Cul3 regulates cytoskeleton protein homeostasis and cell migration during a critical window of brain development." Nature communications1 (2021): 1-22.

• Lu, Yi-Ju, et al. "Arabidopsis calcium-dependent protein kinase 3 regulates actin cytoskeleton organization and immunity." Nature communications1 (2020): 1-12.

• Bojcevski, Jovana et al. “Influence of retinal NMDA receptor activity during autoimmune optic neuritis.” Journal of neurochemistry vol. 153,6 (2020): 693-709. doi:10.1111/jnc.14980

• Chen, Y. et al.  "Coronin 2B regulates dendrite outgrowth by modulating actin dynamics." FEBS Lett, 594: 2975-2987. (2020). https://doi.org/10.1002/1873-3468.13886

• Yang, N. et al. Effects of dexamethasone on remodeling of the hippocampal synaptic filamentous actin cytoskeleton in a model of pilocarpine-induced status epilepticus. Int. J. Med. Sci. 17, 1683–1691 (2020).

• Gupta, R., Kumar, G., Jain, B. P., Chandra, S. & Goswami, S. K. Ectopic expression of 35 kDa and knocking down of 78 kDa SG2NAs induce cytoskeletal reorganization, alter membrane sialylation, and modulate the markers of EMT. Mol. Cell. Biochem. (2020) doi:10.1007/s11010-020-03932-2.

• Musah, A. S. et al. Mechanistic Target of Rapamycin Regulates the Oligodendrocyte Cytoskeleton during Myelination. J. Neurosci. 40, 2993 LP – 3007 (2020).

• Shi, Zengdun et al. “Myocardin and myocardin-related transcription factor-A synergistically mediate actin cytoskeletal-dependent inhibition of liver fibrogenesis.” American journal of physiology. Gastrointestinal and liver physiology vol. 318,3 (2020): G504-G517. doi:10.1152/ajpgi.00302.2019

• Yang, Nuo et al. “Effects of Dexamethasone on Remodeling of the Hippocampal Synaptic Filamentous Actin Cytoskeleton in a Model of Pilocarpine-induced Status Epilepticus.” International journal of medical sciences vol. 17,12 1683-1691. 2 Jul. 2020, doi:10.7150/ijms.44927

• Bojcevski J, Stojic A, Hoffmann DB, Williams SK, Müller A, Diem R, Fairless R. Influence of retinal NMDA receptor activity during autoimmune optic neuritis. J Neurochem. 2020 Jun;153(6):693-709. doi: 10.1111/jnc.14980. Epub 2020Mar 3. PMID: 32031240.

• Kommaddi, Reddy Peera et al. “Aβ mediates F-actin disassembly in dendritic spines leading to cognitive deficits in Alzheimer's disease.” The Journal of neuroscience : the official journal of the Society for Neuroscience vol. 38,5 (2018): 1085-1099. doi:10.1523/JNEUROSCI.2127-17.2017

• Kommaddi RP et al. Aβ mediates F-actin disassembly in dendritic spines leading to cognitive deficits in Alzheimer's disease. J Neurosci. 2018Jan 31;38(5):1085-1099. doi: 10.1523/JNEUROSCI.2127-17.2017. Epub 2017 Dec 15. PMID: 29246925; PMCID: PMC5792472.

• Asahara et al., 2013. Ras-related C3 botulinum toxin substrate 1 (RAC1) regulates glucose-stimulated insulin secretion via modulation of F-actin. Diabetologia. doi: 10.1007/s00125-013-2849-5.

• Ni et al., 2013. The role of RhoA and cytoskeleton in myofibroblast transformation in hyperoxic lung fibrosis. Free Radical Biology and Medicine. doi: http://dx.doi.org/10.1016/j.freeradbiomed.2013.03.012.

• Ramachandran et al., 2013. JunB mediates basal- and TGFb1-induced smooth muscle cell contractility. PLoS ONE. 8(1): e53430.

• Shuang et al., 2013. Destrin deletion enhances the bone loss in hindlimb suspended mice. E. J. Appl. Physiol. 113, 403-410.

• Malenda et al., 2012. Statins Impair Glucose Uptake in Tumor Cells. Neoplasia. 14, 311–323.

• Fan et al., 2012. A role for γS-crystallin in the organization of actin and fiber cell maturation in the mouse lens. FEBS. J. 279, 2892-2904.

• Liu et al., 2012. TLR2 Is a Primary Receptor for Alzheimer's Amyloid β Peptide To Trigger Neuroinflammatory Activation. J. Immunol. 188, 1098-1107.

• Chand et al., 2012. C-terminal region of teneurin-1 co-localizes with dystroglycan and modulates cytoskeletal organization through an extracellular signal-regulated kinase-dependent stathmin- and filamin A-mediated mechanism in hippocampal cells. Neuroscience. 219, 255-270.

• Rapier et al., 2010. The extracellular matrix microtopography drives critical changes in cellular motility and Rho A activity in colon cancer cells. Cancer Cell Int. 10, 24.

• Meeks et al., 2005. Heat shock protein 20-mediated force suppression in forskolin-relaxed swine carotid artery. Am. J. Physiol. 288, C633-C639.

• Zhang et al., 2005. Activation of the Arp2/3 complex by N-WASP is required for actin polymerization and contraction in smooth muscle. Am. J. Physiol. 288, C1145-C1160.

• Chen et al., 2004. Protective effect of phosphatidylinositol 4,5-bisphosphate against cortical filamentous actin loss and insulin resistance induced by sustained exposure of 3T3-L1 adipocytes to insulin. J. Biol. Chem. 279, 39705-39709.

• Tang and Gunst, 2004. The small GTPase Cdc42 regulates actin polymerization and tension development during contractile stimulation of smooth muscle. J. Biol. Chem. 279, 51722-51728.

• Searles et al., 2004. Actin cytoskeleton organization and poststranscriptional regulation of endothelial nitric oxide synthase during cell growth. Circ. Res. 95, 488-495.

• Tu et al., 2003. Migfilin and Mig-2 link focal adhesions to filamin and the actin cytoskeleton and function in cell shape modulation. Cell. 113, 37-47.

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