ホームページ Cytoskeletal Signaling Dynamin inhibitor Dyngo-4a For research use only.

Dyngo-4a

別名：Hydroxy-Dynasore

Dyngo-4a is a potent dynamin inhibitor with IC50 of 0.38 μM, 1.1 μM, and 2.3 μM for DynI (brain), DynI (rec), and DynII (rec), respectively.

Dyngo-4a化学構造

CAS No. 1256493-34-1

サイズ 価格(税別) 在庫状況
10mM (1mL in DMSO) JPY 29500 国内在庫あり
JPY 22000 国内在庫あり
JPY 52000 国内在庫あり
JPY 104500 国内在庫あり
JPY 295500 国内在庫なし(納期7~10日)

代表番号: 045-509-1970|電子メール：[email protected]
よく尋ねられる質問

文献中Selleckの製品使用例（26）

カスタマーフィードバック3例

製品安全説明書

現在のバッチを見る： S716302 DMSO] 68 mg/mL] false] Water] Insoluble] false] Ethanol] Insoluble] false 純度： 98.33%
98.33

Dyngo-4a関連製品

Dynamin阻害剤の選択性比較

生物活性

製品説明 Dyngo-4a is a potent dynamin inhibitor with IC50 of 0.38 μM, 1.1 μM, and 2.3 μM for DynI (brain), DynI (rec), and DynII (rec), respectively.
Targets
DynI (brain) [1] DynI (rec) [1] DynII (rec) [1]
0.38 μM 1.1 μM 2.3 μM
In Vitro
In vitro Dyngo-4a inhibits dynamin-dependent endocytosis of transferrin in multiple cell types with IC₅₀ of 5.7 μM, and reduces synaptic vesicle endocytosis and activity-dependent bulk endocytosis in cultured neurons and synaptosomes. [1] In motor nerve terminals and cultured hippocampal neurons, Dyngo-4a blocks Alexa Fluor 488-BoNT/A-Hc internalization. [2] In Drosophila S2R+ cells, Dyngo-4a causes a decrease in the level of Armadillo/β-catenin. [3]
Kinase Assay Dynamin GTPase assay
Dynamin I activity is measured in its SAI activity state or is stimulated by three different methods. As each stimulus activates dynamin to different extents, each assay requires different dynamin concentrations. First, maximal dynamin activity is stimulated by sonicated PS liposomes. Purified dynamin I (10–20 nM, diluted in: 6 mM Tris–HCl, 20 mM NaCl and 0.01% Tween 80, pH 7.4) is incubated in 96-well plates in GTPase buffer (5 mM Tris–HCl, 10 mM NaCl, 2 mM Mg2+, 0.05% Tween 80, pH 7.4, 1 µg/mL leupeptin and 0.1 mM PMSF) and GTP 0.3 mM in the presence of test compound for 30 min at 37°C in a final assay volume of 150 μL. Reactions are terminated with 10 μL of 0.5 M ethylenediaminetetraacetic acid (EDTA) pH 7.4 and Malachite Green solution (40 μL: 2% w/v ammonium molybdate tetrahydrate, 0.15% w/v malachite green and 4 M HCl) is added for 5 min. Second, dynamin (20 nM) is stimulated by 10 µg/mL of taxol-stabilized preformed bovine brain microtubules using the same protocol. Third, dynamin I (50 nM) is stimulated by 1 μM of recombinant grb2, a SH3-containing protein that stimulates dynamin about 5–10 times less efficiently than liposomes or microtubules. Finally, dynamin (500 nM) SAI activity is measured using high concentrations of dynamin, which promote its cooperative self-assembly into rings (but not helices). The final DMSO concentration in the GTPase or endocytosis assays is at most 3.3 or 1%, respectively, but typically is at 1%. The GTPase assay for dynamin I is unaffected by DMSO up to 3.3%. Compounds are dissolved as 30 mM stocks in 100% DMSO. These stock solutions can be stored at −20°C for several months. Compounds are subsequently diluted into solutions of 50% DMSO made up in 20 mM Tris–HCl pH 7.4 and diluted again into the final assay. For analysis of the kinetics of 4a inhibition, dynamin I at a final concentration of 17 nM is incubated with GTPase buffer containing PS (2 µg/mL) and varying amounts of GTP (50–250 μM) in the presence of 4a at a concentration range between 0.5 and 6 μM. The reaction is stopped after 30 min by addition of EDTA (0.5 mM, pH 7.4). Curves are generated using the Michaelis–Menten equation v = Vmax[S]/(Km + [S]), where S is the GTP substrate. After the Vmax and Km values were determined, the data were transformed using the Lineweaver–Burke equation, 1/v = 1/Vmax + (Km/Vmax)(1/[S]). Assay conditions are based on the dynamin I assay but contained modifications. Recombinant dynamin II is used at 50 nM, stimulated by 10 µg/mL PS. The GTPase reaction is allowed to occur for 90 min at 37°C before termination.
細胞実験 細胞株 αT3–1 or LβT2 cells
濃度 30 μM
反応時間 30 minutes
実験の流れ

αT3-1 or LβT2 cells (2 × 106) were grown overnight in a 6-well culture dish and then serum starved for 2–4 hours. Cells were pretreated with either vehicle (0), dynasore (80μM), or dyngo (30μM) for 30 minutes. For dose-response studies, indicated doses of dynasore were used for a 30-minute pretreatment. After pretreatment, cells were treated with 0 or GnRHa (10 nM) for 10 minutes. Cells were then washed twice in PBS, lysed in radio-immunoprecipitation assay (RIPA) buffer and subjected to SDS-PAGE (acrylamide:bis-acrylamide ratio of 29:1) and electroblotted to polyvinylidene difluoride membranes.

化学情報

分子量 338.31 化学式

C18H14N2O5
CAS No. 1256493-34-1 SDF Download Dyngo-4a SDFをダウンロードする
Smiles C1=CC=C2C=C(C(=CC2=C1)C(=O)NN=CC3=CC(=C(C=C3O)O)O)O
保管

In vitro
Batch:

DMSO : 68 mg/mL ( (200.99 mM)； 吸湿したDMSOは溶解度を減少させます。新しいDMSOをご使用ください。)

Water : Insoluble

Ethanol : Insoluble

モル濃度計算器

in vivo
Batch:

Add solvents to the product individually and in order.

投与溶液組成計算機

実験計算

モル濃度計算器

質量 濃度 体積 分子量

投与溶液組成計算機(クリア溶液)

ステップ１：実験データを入力してください。（実験操作によるロスを考慮し、動物数を1匹分多くして計算・調製することを推奨します）

mg/kg g μL 匹

ステップ２：投与溶媒の組成を入力してください。（ロット毎に適した溶解組成が異なる場合があります。詳細については弊社までお問い合わせください）

% DMSO % % Tween 80 % ddH2O
%DMSO %

計算結果：

投与溶媒濃度： mg/ml;

DMSOストック溶液調製方法： mg 試薬を μL DMSOに溶解する（濃度 mg/mL, 注：濃度が当該ロットのDMSO溶解度を超える場合はご連絡ください。 )

投与溶媒調製方法：Take μL DMSOストック溶液に μL PEG300，を加え、完全溶解後μL Tween 80，を加えて完全溶解させた後 μL ddH2O，を加え完全に溶解させます。

投与溶媒調製方法：μL DMSOストック溶液に μL Corn oil，を加え、完全溶解。

注意：１.ストック溶液に沈殿、混濁などがないことをご確認ください；
２.順番通りに溶剤を加えてください。次のステップに進む前に溶液に沈殿、混濁などがないことを確認してから加えてください。ボルテックス、ソニケーション、水浴加熱など物理的な方法で溶解を早めることは可能です。

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