Forskolin 佛司可林/毛喉素
產品標簽
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）（一種12-跨膜域酶）產生，AC催化ATP轉化生成3′,5′-cAMP和焦磷酸鹽。cAMP是一種重要的信號載體，在細胞對激素，神經遞體和其他細胞外信號的恰當的生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信號通過靶向信號組分到亞細胞區室和組裝信號復合物來獲得。
產品描述
佛司可林（Forskolin，CAS NO.：66575-29-9）是一種來源于印度彩葉植物毛喉鞘蕊花（Coleus forskohlii）的天然生成二萜。通過作用于催化亞基直接激活腺苷酸環化酶（AC），引起細胞內cAMP水平的提高[1-2]。在穩定轉染且表達I型腺苷酸環化酶（AC1）的Sf9細胞，Forskolin與膜上的AC1結合（IC50=41nM）并活化其功能（EC50=0.5 μM，評估從ATP生成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，同時影響細胞質鈣離子水平和收縮成分的鈣離子敏感性[4]。Forskolin能夠與葡萄糖轉運體和某些離子通道相互作用，可用來檢測腺苷酸環化酶的表達、調控和G蛋白信號通路[5]。Forskolin用于幾種分化方案中以增強神經元分化能力[6-7]。
我司提供兩種形式的佛司可林（Forskolin）：
一種是溶于DMSO的50mM儲存液形式，對應貨號是：MZ2301-50UL（~1mg），MZ2301-250UL（~5mg），直接用培養基稀釋到所需工作濃度即可。操作簡單方便。
一種是凍干粉形式，對應貨號是：MZ2301-50MG，MZ2301-100MG。需要先用DMSO或無水乙醇配制成儲存液，再用培養基稀釋到所需工作濃度使用。
Forskolin通常用在各種類型的完整細胞或組織培養物中來提高cAMP水平，常用工作濃度為10-50μM。具體的最佳工作濃度請參考文獻，或根據自身實驗體系，培養細胞和組織類型，通過實驗進行摸索和優化。
產品應用（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)
作用機制：Jurkat細胞能生成IL-2。正常生長情況，Jurkat細胞幾乎沒有或不生成IL-2。PMA，通過刺激PKC能活化T細胞并刺激低水平的IL-2生成。當Jurkat細胞經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。PHA本身能夠激活低水平的T細胞活化和IL-2生成，通過非特異性結合到細胞表面受體復合物。T淋巴細胞中，Forskolin激活腺苷酸環化酶會抑制IL-2合成。Forskolin通過間接干擾磷脂酶C活化來發揮這一功能。Forskolin激活腺苷酸環化酶，產生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
5）分子式：C22H34O7
6）分子量：410.50 g/mol
7）純度：>99%
8）外觀：白色結晶性粉末
9）化學結構式：
保存與運輸方法
保存：凍干粉-20oC避光保存，至少3年有效；DMSO儲存液-20oC避光保存至少6個月有效；
運輸：冰袋運輸。
使用方法【源自文獻，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 體內研究： 動物模型（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。小規格提供50μl（50mM in DMSO），按照10-50μM的工作濃度來算，可配制50ml-250ml工作液。
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產品名稱 |
產品編號 |
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 |
注意事項
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，請穿實驗服并戴一次性手套操作。
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】
上海懋康生物科技有限公司是一家涉足于生命科學和生物技術領域研究的試劑、儀器和實驗室消耗品與實驗服務工作，主要從事細胞生物學、植物學、分子生物學、免疫學、生物化學、蛋白組學。生物制藥與診斷試劑研發生產等領域。 本公司秉承“以人為本，以誠為信、合同守信”的經營理念。堅持"品質保障"的原則為廣大客戶提供優質產品。
