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        当前位置:首页> 新闻资讯> 新品发布 >MKbio Forskolin 佛司可林/毛喉素(最经典的腺苷酸环化酶活化剂)
        MKbio Forskolin 佛司可林/毛喉素(最经典的腺苷酸环化酶活化剂)
        时间:2018-01-25     作者:懋康原创     文章来源:懋康生物    

        Forskolin 佛司可林/毛喉素


        产品标签

        Forskolin (Fsk) 佛司可林;Adenylate cyclase (AC) 腺苷酸环化酶;cAMP 环磷酸腺苷;Neuron Differentiation 神经元分化;CAS NO:66575-29-9;


        景描述

        环磷酸腺苷(cAMP)由腺苷酸环化酶(Adenylate cyclase , AC)(一种12-跨膜域酶)产生,AC催化ATP转化生成3′,5′-cAMP和焦磷酸盐。cAMP是一种重要的信号载体,在细胞对激素,神经递体和其他细胞外信号的恰当的生理反应中是不可或缺的。环磷酸腺苷(cAMP)由AC合成且经磷酸二酯酶(phosphodiesterases, PDEs)降解。局部cAMP信号通过靶向信号组分到亚细胞区室和组装信号复合物来获得。 


        产品描述

        佛司可林(Forskolin,CAS NO.:66575-29-9)是一种来源于印度彩叶植物毛喉鞘蕊花(Coleus forskohlii)的天然生成二萜。通过作用于催化亚基直接激活腺苷酸环化酶(AC),引起细胞内cAMP水平的提高[1-2]。在稳定转染且表达I型腺苷酸环化酶(AC1)的Sf9细胞,Forskolin与膜上的AC1结合(IC50=41nM)并活化其功能(EC50=0.5 μM,评估从ATP生成cAMP的激活实验)[3]。Forskolin以浓度依赖性的方式发挥平滑肌松弛剂和血管扩张剂的功能,同时影响细胞质钙离子水平和收缩成分的钙离子敏感性[4]。Forskolin能够与葡萄糖转运体和某些离子通道相互作用,可用来检测腺苷酸环化酶的表达、调控和G蛋白信号通路[5]。Forskolin用于几种分化方案中以增强神经元分化能力[6-7]。


        我司提供两种形式的佛司可林(Forskolin):

        一种是溶于DMSO的50mM储存液形式,对应货号是:MZ2301-50UL(~1mg),MZ2301-250UL(~5mg),直接用培养基稀释到所需工作浓度即可。操作简单方便。

        一种是冻干粉形式,对应货号是:MZ2301-50MG,MZ2301-100MG。需要先用DMSO或无水乙醇配制成储存液,再用培养基稀释到所需工作浓度使用。


        Forskolin通常用在各种类型的完整细胞或组织培养物中来提高cAMP水平,常用工作浓度为10-50μM。具体的最佳工作浓度请参考文献,或根据自身实验体系,培养细胞和组织类型,通过实验进行摸索和优化。



        产品应用(T淋巴细胞使用Forskolin抑制IL-2合成的应用示例)

        细胞类型:Jurkat cells (淋巴细胞样T细胞系)

        作用机制:Jurkat细胞能生成IL-2。正常生长情况,Jurkat细胞几乎没有或不生成IL-2。PMA,通过刺激PKC能活化T细胞并刺激低水平的IL-2生成。当Jurkat细胞经PMA和共刺激剂比如植物血凝素PHA处理,IL-2的生成明显得到加强。PHA本身能够激活低水平的T细胞活化和IL-2生成,通过非特异性结合到细胞表面受体复合物。T淋巴细胞中,Forskolin激活腺苷酸环化酶会抑制IL-2合成。Forskolin通过间接干扰磷脂酶C活化来发挥这一功能。Forskolin激活腺苷酸环化酶,产生cAMP,从而激活PKA。PKA能磷酸化PLC的丝氨酸残基,从而间接抑制IL-2生成。

        反应步骤:

        2.5 mL of Jurkat cells (1×106cells/mL) and 2.5 mL fresh culture medium (RPMI-1640 with 10% fetal calf serum containing 10 mL/L penicillin-streptomycin) were added to 25 cm2culture bottles. The following additions were made in duplicate.

        a. Control - no additions

        b. 1 μg/mL PHA + 10 ng/mL PMA [Add 10 μL of PHA stock solution (0.5 mg/mL PHA in filter-sterilized PBS) and 5 μL of PMA stock solution (10 μg/mL PMA in DMSO)]

        c. 0.2 mM Forskolin + 1 μg/mL PHA + 10 ng/mL PMA [Add 10 μL of PHA stock solution, 5 μL of PMA stock solution, and 10 μL of forskolin stock solution (100 mM forskolin in DMSO)

        After mixing well, the bottles were incubated at 37 °C for 24 hours. After centrifugation, the clarified broth was then tested for IL-2 production by ELISA assay. IL-2 production in the test cultures containing 0.2 mM forskolin was inhibited ≥50% compared to the test cultures containing only PHA and PMA. 


        产品特性

        1)CAS NO:66575-29-9

        2)化学名:5-(acetyloxy)-3-ethenyldodecahydro-6,10,10b-trihydroxy-3,4a,7,7,10a-pentamethyl-(3R,4aR,5S,6S, 6aS,10S,10aR,10bS)-1H-naphtho[2,1-b]pyran-1-one

        3)同义名:Coleonol洋紫苏醇;NSC 375489;NSC 357088;L 75-1362B;HL 362;

        4)溶解性:溶于乙醇(~15 mg/ml),DMSO(~30 mg/ml),DMF(~30 mg/ml),几乎不溶于水

        5)分子式:C22H34O7

        6)分子量:410.50 g/mol

        7)纯度:>99%

        8)外观:白色结晶性粉末

        9)化学结构式:


        保存与运输方法

        保存:冻干粉-20oC避光保存,至少3年有效;DMSO储存液-20oC避光保存至少6个月有效;

        运输:冰袋运输。


        使用方法【源自文献,仅作参考】

        文献1,Rodriguez G et al.Forskolin-inducible cAMP pathway negatively regulates T-cell proliferation by uncoupling the interleukin-2 receptor complex. J Biol Chem. 2013 Mar 8;288(10):7137-46.PMID: 23341462

        体外研究:

        细胞类型(Cell type):Human leukemic cell line MT-2 cells

        药物配制(Preparation):Forskolinwas dissolved in DMSO to make stock solution.

        实验方法(Assay):MT-2 cells were seeded into 96-well plates at 5 × 104 cells per well. Cells were then pretreated for1 h with 1% DMSO (vehicle) or Fsk at 1, 5, 10, 25, 50, and 100 μM concentrations. The cells were stimulated with IL-2 as above and cultured for an additional 20 h at 37 °C. Control cells were treated with 1% DMSO for 20 h. During the final 4 h of incubation, the cells were pulsed with [3H]thymidine at a concentration of 0.5 μCi/200 μl. Cells were harvested onto fiberglass filters and analyzed using liquid scintillation counting.Fsk treatment inhibited the proliferation of MT-2 cells in a dose-dependent manner with an IC50 equal to~5 μM Fsk. 

        文献2,Matsumiya W et al. Forskolin modifies retinal vascular development in Mrp4-knockout mice. Invest Ophthalmol Vis Sci. 2012 Dec 7;53(13):8029-35. PMID:23154460

        体内研究:

        动物模型(Animal Model):C57BL/6J mice and Mrp4-knockout mice

        药物配制(Preparation):Forskolin was dissolved in dimethyl sulfoxide (DMSO) and then diluted.

        注射剂量(Dosages):Forskolin was dissolved in DMSO and injected intraperitoneally into neonatal mice at postnatal days 4 (P4) (1.0 μg/50 μL (0.3 mg/kg))and 5 (P5)(1.5 μg/50 μL (0.5 mg/kg)). Mice injected with DMSO served as the controls. The treated mice were euthanized at P6, and their retinas were isolated for whole-mount immunohistochemistry (IHC)

        给药途径(Administration):Intraperitoneally (i.p.) Injection


        订购信息:进口原料(广泛应用于全球各大实验室,纯度>99%),质量保证。小规格提供50μl(50mM in DMSO),按照10-50μM的工作浓度来算,可配制50ml-250ml工作液。 

        产品名称

        产品编号

        规格             

        CAS NO.       

        价格(元)      

        Forskolin (50mM in DMSO) 佛司可林    

        MZ2301-50UL        

        50μl

        66575-29-9

        150

        Forskolin (50mM in DMSO) 佛司可林

        MZ2301-250UL

        5×50μl

        66575-29-9

        520

        Forskolin (Powder) 佛司可林

        MZ2301-50MG

        50mg

        66575-29-9

        1480

        Forskolin (Powder) 佛司可林

        MZ2301-100MG

        100mg

        66575-29-9

        2540


        注意事项

        1)本品不是临床药物,只能用于科研用途,不能用于诊断或临床用途。

        2)为了您的安全和健康,请穿实验服并戴一次性手套操作。


        参考文献

        [1] Barber, R. and Goka, T.J. "Adenylate cyclase activity as a function of forskolin concentration." J. Cyclic Nucleotide Protein Phosphor. Res. 10: 23-29 (1985).

        [2] Laurenza, A., et al. "Forskolin: a specific stimulator of adenylyl cyclase or a diterpene with multiple sites of action?" Trends Pharm. Sci. 10: 442-447 (1989).

        [3] Robbins JD et al. Forskolin carbamates: binding and activation studies with type I adenylyl cyclase. J Med Chem. 1996 Jul 5;39(14):2745-52.

        [4] Abe, A. and Karaki, H.J. "Effect of forskolin on cytosolic Ca++ level and contraction in vascular smooth muscle." J. Pharmacol. Exp. Ther. 249: 895-900 (1989).

        [5] Insel PA et al. Forskolin as a tool for examining adenylyl cyclase expression, regulation, and G protein signaling. Cell Mol Neurobiol. 2003 Jun;23(3):305-14.

        [6] Palmer TD et al. The adult rat hippocampus contains primordial neural stem cells.Mol Cell Neurosci. 1997;8(6):389-404.

        [7] Son H et al. Pairing of forskolin and KCl increases differentiation of immortalized hippocampal neurons in a CREB Serine 133 phosphorylation-dependent and extracellular-regulated protein kinase-independent manner. Neurosci Lett. 2001 Jul 27;308(1):37-40.


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         — —Written/Edited by V. Shallan【版权归MKBio懋康所有】

         

         

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