Forskolin 佛司可林/毛喉素
産品标簽
Forskolin (Fsk) 佛司可林；Adenylate cyclase (AC) 腺苷酸環化酶；cAMP 環磷酸腺苷；Neuron Differentiation 神經元分化；CAS NO：66575-29-9；
背景描述
環磷酸腺苷（cAMP）由腺苷酸環化酶（Adenylate cyclase , AC）（一(yī)種12-跨膜域酶）産生(shēng)，AC催化ATP轉化生(shēng)成3′,5′-cAMP和焦磷酸鹽。cAMP是一(yī)種重要的信号載體，在細胞對激素，神經遞體和其他細胞外信号的恰當的生(shēng)理反應中是不可或缺的。環磷酸腺苷（cAMP）由AC合成且經磷酸二酯酶（phosphodiesterases, PDEs）降解。局部cAMP信号通(tōng)過靶向信号組分到(dào)亞細胞區室和組裝信号複合物(wù)來獲得。
産品描述
佛司可林（Forskolin，CAS NO.：66575-29-9）是一(yī)種來源于印度彩葉植物(wù)毛喉鞘蕊花（Coleus forskohlii）的天然生(shēng)成二萜。通(tōng)過作用于催化亞基直接激活腺苷酸環化酶（AC），引起細胞内cAMP水(shuǐ)平的提高(gāo)[1-2]。在穩定轉染且表達I型腺苷酸環化酶（AC1）的Sf9細胞，Forskolin與膜上(shàng)的AC1結合（IC50=41nM）并活化其功能(néng)（EC50=0.5 μM，評估從(cóng)ATP生(shēng)成cAMP的激活實驗）[3]。Forskolin以濃度依賴性的方式發揮平滑肌松弛劑和血管擴張劑的功能(néng)，同時影響細胞質鈣離子水(shuǐ)平和收縮成分的鈣離子敏感性[4]。Forskolin能(néng)夠與葡萄糖轉運體和某些離子通(tōng)道相(xiàng)互作用，可用來檢測腺苷酸環化酶的表達、調控和G蛋白(bái)信号通(tōng)路(lù)[5]。Forskolin用于幾種分化方案中以增強神經元分化能(néng)力[6-7]。
我司提供兩種形式的佛司可林（Forskolin）：
一(yī)種是溶于DMSO的50mM儲存液形式，對應貨号是：MZ2301-50UL（~1mg），MZ2301-250UL（~5mg），直接用培養基稀釋到(dào)所需工(gōng)作濃度即可。操作簡單方便。
一(yī)種是凍幹粉形式，對應貨号是：MZ2301-50MG，MZ2301-100MG。需要先用DMSO或無水(shuǐ)乙醇配制成儲存液，再用培養基稀釋到(dào)所需工(gōng)作濃度使用。
Forskolin通(tōng)常用在各種類型的完整細胞或組織培養物(wù)中來提高(gāo)cAMP水(shuǐ)平，常用工(gōng)作濃度為(wèi)10-50μM。具體的最佳工(gōng)作濃度請參考文獻，或根據自(zì)身實驗體系，培養細胞和組織類型，通(tōng)過實驗進行摸索和優化。
産品應用（T淋巴細胞使用Forskolin抑制IL-2合成的應用示例）
細胞類型：Jurkat cells (淋巴細胞樣T細胞系)
作用機(jī)制：Jurkat細胞能(néng)生(shēng)成IL-2。正常生(shēng)長(cháng)情況，Jurkat細胞幾乎沒有或不生(shēng)成IL-2。PMA，通(tōng)過刺激PKC能(néng)活化T細胞并刺激低(dī)水(shuǐ)平的IL-2生(shēng)成。當Jurkat細胞經PMA和共刺激劑比如植物(wù)血凝素PHA處理，IL-2的生(shēng)成明顯得到(dào)加強。PHA本身能(néng)夠激活低(dī)水(shuǐ)平的T細胞活化和IL-2生(shēng)成，通(tōng)過非特異性結合到(dào)細胞表面受體複合物(wù)。T淋巴細胞中，Forskolin激活腺苷酸環化酶會(huì)抑制IL-2合成。Forskolin通(tōng)過間接幹擾磷脂酶C活化來發揮這一(yī)功能(néng)。Forskolin激活腺苷酸環化酶，産生(shēng)cAMP，從(cóng)而激活PKA。PKA能(néng)磷酸化PLC的絲氨酸殘基，從(cóng)而間接抑制IL-2生(shēng)成。
反應步驟：
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.
産品特性
1）CAS NO：66575-29-9
2）化學名：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(shuǐ)
5）分子式：C22H34O7
6）分子量：410.50 g/mol
7）純度：>99%
8）外觀：白(bái)色結晶性粉末
9）化學結構式：
保存與運輸方法
保存：凍幹粉-20ºC避光(guāng)保存，至少3年(nián)有效；DMSO儲存液-20ºC避光(guāng)保存至少6個(gè)月(yuè)有效；
運輸：冰袋運輸。
使用方法【源自(zì)文獻，僅作參考】
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文獻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： 細胞類型（Cell type）：Human leukemic cell line MT-2 cells 藥物(wù)配制（Preparation）：Forskolinwas dissolved in DMSO to make stock solution. 實驗方法（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. |
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文獻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 體内研究： 動物(wù)模型（Animal Model）：C57BL/6J mice and Mrp4-knockout mice 藥物(wù)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 |
訂購信息：進口原料（廣泛應用于全球各大實驗室，純度＞99%），質量保證。小(xiǎo)規格提供50μl（50mM in DMSO），按照(zhào)10-50μM的工(gōng)作濃度來算(suàn)，可配制50ml-250ml工(gōng)作液。
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産品名稱 |
産品編号 |
規格 |
CAS NO. |
價格（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(shì)項
1）本品不是臨床藥物(wù)，隻能(néng)用于科研用途，不能(néng)用于診斷或臨床用途。
2）為(wèi)了您的安全和健康，請穿實驗服并戴一(yī)次性手套操作。
參考文獻
[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【版權歸MKBio懋康所有】
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