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| 上海懋康生物科技有限公司
主營(yíng)產(chǎn)品： 硫酸諾爾斯菌素,D-熒光素,雷帕霉素 |
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2018-3-7 閱讀(2260)
Forskolin 佛司可林/毛喉素
產(chǎn)品標(biāo)簽
Forskolin (Fsk) 佛司可林；Adenylate cyclase (AC) 腺苷酸環(huán)化酶；cAMP 環(huán)磷酸腺苷；Neuron Differentiation 神經(jīng)元分化；CAS NO：66575-29-9；
背景描述
環(huán)磷酸腺苷（cAMP）由腺苷酸環(huán)化酶（Adenylate cyclase , AC）（一種12-跨膜域酶）產(chǎn)生，AC催化ATP轉(zhuǎn)化生成3′,5′-cAMP和焦磷酸鹽。cAMP是一種重要的信號(hào)載體，在細(xì)胞對(duì)激素，神經(jīng)遞體和其他細(xì)胞外信號(hào)的恰當(dāng)?shù)纳矸磻?yīng)中是*的。環(huán)磷酸腺苷（cAMP）由AC合成且經(jīng)磷酸二酯酶（phosphodiesterases, PDEs）降解。局部cAMP信號(hào)通過(guò)靶向信號(hào)組分到亞細(xì)胞區(qū)室和組裝信號(hào)復(fù)合物來(lái)獲得。
產(chǎn)品描述
佛司可林（Forskolin，CAS NO.：66575-29-9）是一種來(lái)源于印度彩葉植物毛喉鞘蕊花（Coleus forskohlii）的天然生成二萜。通過(guò)作用于催化亞基直接激活腺苷酸環(huán)化酶（AC），引起細(xì)胞內(nèi)cAMP水平的提高[1-2]。在穩(wěn)定轉(zhuǎn)染且表達(dá)I型腺苷酸環(huán)化酶（AC1）的Sf9細(xì)胞，F(xiàn)orskolin與膜上的AC1結(jié)合（IC50=41nM）并活化其功能（EC50=0.5 μM，評(píng)估從ATP生成cAMP的激活實(shí)驗(yàn)）[3]。Forskolin以濃度依賴(lài)性的方式發(fā)揮平滑肌松弛劑和血管擴(kuò)張劑的功能，同時(shí)影響細(xì)胞質(zhì)鈣離子水平和收縮成分的鈣離子敏感性[4]。Forskolin能夠與葡萄糖轉(zhuǎn)運(yùn)體和某些離子通道相互作用，可用來(lái)檢測(cè)腺苷酸環(huán)化酶的表達(dá)、調(diào)控和G蛋白信號(hào)通路[5]。Forskolin用于幾種分化方案中以增強(qiáng)神經(jīng)元分化能力[6-7]。
我司提供兩種形式的佛司可林（Forskolin）：
一種是溶于DMSO的50mM儲(chǔ)存液形式，對(duì)應(yīng)貨號(hào)是：MZ2301-50UL（~1mg），MZ2301-250UL（~5mg），直接用培養(yǎng)基稀釋到所需工作濃度即可。操作簡(jiǎn)單方便。
一種是凍干粉形式，對(duì)應(yīng)貨號(hào)是：MZ2301-50MG，MZ2301-100MG。需要先用DMSO或無(wú)水乙醇配制成儲(chǔ)存液，再用培養(yǎng)基稀釋到所需工作濃度使用。
Forskolin通常用在各種類(lèi)型的完整細(xì)胞或組織培養(yǎng)物中來(lái)提高cAMP水平，常用工作濃度為10-50μM。具體的zui加工作濃度請(qǐng)參考文獻(xiàn)，或根據(jù)自身實(shí)驗(yàn)體系，培養(yǎng)細(xì)胞和組織類(lèi)型，通過(guò)實(shí)驗(yàn)進(jìn)行摸索和優(yōu)化。
產(chǎn)品應(yīng)用（T淋巴細(xì)胞使用Forskolin抑制IL-2合成的應(yīng)用示例）
細(xì)胞類(lèi)型：Jurkat cells (淋巴細(xì)胞樣T細(xì)胞系)
作用機(jī)制：Jurkat細(xì)胞能生成IL-2。正常生長(zhǎng)情況，Jurkat細(xì)胞幾乎沒(méi)有或不生成IL-2。PMA，通過(guò)刺激PKC能活化T細(xì)胞并刺激低水平的IL-2生成。當(dāng)Jurkat細(xì)胞經(jīng)PMA和共刺激劑比如植物血凝素PHA處理，IL-2的生成明顯得到加強(qiáng)。PHA本身能夠激活低水平的T細(xì)胞活化和IL-2生成，通過(guò)非特異性結(jié)合到細(xì)胞表面受體復(fù)合物。T淋巴細(xì)胞中，F(xiàn)orskolin激活腺苷酸環(huán)化酶會(huì)抑制IL-2合成。Forskolin通過(guò)間接干擾磷脂酶C活化來(lái)發(fā)揮這一功能。Forskolin激活腺苷酸環(huán)化酶，產(chǎn)生cAMP，從而激活PKA。PKA能磷酸化PLC的絲氨酸殘基，從而間接抑制IL-2生成。
反應(yīng)步驟：
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.
產(chǎn)品特性
1）CAS NO：66575-29-9
2）化學(xué)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）同義名：Coleonol洋紫蘇醇；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），幾乎不溶于水
5）分子式：C22H34O7
6）分子量：410.50 g/mol
7）純度：>99%
8）外觀：白色結(jié)晶性粉末
9）化學(xué)結(jié)構(gòu)式：
使用方法【源自文獻(xiàn)，僅作參考】
文獻(xiàn)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 體外研究： 細(xì)胞類(lèi)型（Cell type）：Human leukemic cell line MT-2 cells 藥物配制（Preparation）：Forskolinwas dissolved in DMSO to make stock solution. 實(shí)驗(yàn)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. |
文獻(xiàn)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 體內(nèi)研究： 動(dòng)物模型（Animal Model）：C57BL/6J mice and Mrp4-knockout mice 藥物配制（Preparation）：Forskolin was dissolved in dimethyl sulfoxide (DMSO) and then diluted. 注射劑量（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) 給藥途徑（Administration）：Intraperitoneally (i.p.) Injection |
訂購(gòu)信息：進(jìn)口原料（廣泛應(yīng)用于各大實(shí)驗(yàn)室，純度＞99%），質(zhì)量保證。小規(guī)格提供50μl（50mM in DMSO），按照10-50μM的工作濃度來(lái)算，可配制50ml-250ml工作液。
產(chǎn)品名稱(chēng) | 產(chǎn)品編號(hào) | 規(guī)格 | CAS NO. | 價(jià)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 |
注意事項(xiàng)
1）本品不是臨床藥物，只能用于科研用途，不能用于診斷或臨床用途。
2）為了您的安全和健康，請(qǐng)穿實(shí)驗(yàn)服并戴一次性手套操作。
參考文獻(xiàn)
[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.
相關(guān)產(chǎn)品
貨號(hào) | 名稱(chēng) | 規(guī)格 |
MZ2301-50UL | Forskolin (50mM in DMSO) 佛司可林 | 50μl |
MZ2301-50MG | Forskolin (Powder) 佛司可林 | 50mg |
MZ2401-100UL | PMA (TPA, 5mg/ml) 佛波肉荳蔻醋酸（5mg/ml） | 100μl |
MZ2402-5MG | PMA (TPA, Powder) 佛波肉荳蔻醋酸（粉末，超純） | 5mg |
MZ2403-1MG | 4α-PMA (4α-TPA) 4α-佛波肉荳蔻醋酸 | 1mg |
MP6303-5MG | Phytohemagglutinin-M (PHA-M) 植物血凝素-M（菜豆） | 5mg |
MP6305-5MG | Phytohemagglutinin-P (PHA-P) 植物血凝素-P（菜豆） | 5mg |
— —Written/Edited by V. Shallan【版權(quán)歸MKBio懋康所有】
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