Dyngo-4a

Catalog No.S7163

Dyngo-4a Chemical Structure

Molecular Weight(MW): 338.31

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.

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Cited by 4 Publications

3 Customer Reviews

  • Immunofluorescence analysis of EGFR distribution in HCC827 and H1650 cells treated with dynasore, dyngo-4a, or control reagents. Images were captured using a fluorescent microscope (Olympus BX63, 40X objective). Scale bar = 10 μm.

    Cell Commun Signal, 2018, 16(1):40. Dyngo-4a purchased from Selleck.

    A) Purified splenic CD4+ T-cells were activated in the presence or absence of the indicated concentrations of Dyngo-4a. Nuclear and cytoplasmic extracts were then prepared. Western blots were probed with αICN (110kDa), αNucleolin (110kDa) and αActin (42kDa). Densitometry values for ICN bands in nuclear and cytoplasmic extracts were normalized to those of nucleolin and actin bands, respectively. Quantification of 2 independent experiments is shown in bar graph. Error bars represent ±SEM.

    J Immunol, 2018, 200(3):997-1007. Dyngo-4a purchased from Selleck.

  • cultured HCC827 and H1650 cells were treated with 20 μM of dyngo-4a for indicated times and lysed. The expression levels of EGFR, pAKT, and pMEK were examined by immunoblotting. GAPDH was probed to confirm equal loading. Dyngo-4a did not repress the phosphorylation of AKT or MEK in neither HCC827 nor H1650 cells with various treatment times under normal culture condition.

    Int J Biochem Cell Biol, 2018, 105:1-12. Dyngo-4a purchased from Selleck.

Purity & Quality Control

Choose Selective Dynamin Inhibitors

Biological Activity

Description 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

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]

Protocol

Kinase Assay:

[1]

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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.
Cell Research:

[5]

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  • Cell lines: αT3–1 or LβT2 cells
  • Concentrations: 30 μM
  • Incubation Time: 30 minutes
  • Method:

    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.


    (Only for Reference)
Animal Research:

[4]

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  • Animal Models: Female CD-1 mice
  • Formulation: 1/9 NMP/PEG300 in PBS
  • Dosages: 30 mg/kg
  • Administration: i.p.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 67 mg/mL (198.04 mM)
Ethanol 1 mg/mL warmed (2.95 mM)
Water Insoluble

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 338.31
Formula

C18H14N2O5

CAS No. 1256493-34-1
Storage powder
in solvent
Synonyms N/A

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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID