EG00229 is a small molecular inhibitor of VEGF-A interaction with NRP-1.29 (link)–31 (link) For cell culture studies, EG00229 (R&D Systems, #4931) was dissolved in DMSO to produce a 100 mM (50 mg/ml) stock. For in vivo studies, EG00229 was diluted to 2 mM (1 mg/ml) in 20% captisol, a non-toxic delivery vehicle.57 (link) In brief, 40% captisol (RC-0C7-020, CyDex Pharmaceuticals, Lawrence, KS, USA) was prepared in sterile water by stirring overnight at 25°C followed by filter sterilization. The 40% captisol solution (50 ml) was diluted 1:1 with sterile water and supplemented with 0.5 ml of 1N acetic acid to create 20% captisol solution. The 50 mg/ml EG00229 stock was diluted 1:50 into 20% captisol solution and 0.2 ml was injected intraperitoneally three times per week (M, W, F) to achieve a final level of 10 mg/kg body weight.
Eg00229
Manufactured by R&D Systems
Sourced in Germany
EG00229 is a laboratory equipment used for analysis and measurement purposes. It is designed to perform specific tasks related to research and development. The core function of this product is to provide accurate and reliable data, but a detailed description while maintaining an unbiased and factual approach is not available.
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3 protocols using eg00229
1
EG00229: VEGF-A Inhibitor in Cell and Animal Studies
2
Regulation of VEGF-A Signaling Pathways
3
Cellular Uptake of Nanoparticles Modulated by RGE and NRP-1 Inhibitor
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Cellular uptake study was performed with the U87MG cell line. Cells were seeded in a glass-bottom dish or 6-well plates at a density of 3 × 104 cells/mL and cultured for 24 h. The cells were divided into 3 groups: RGE-modified NPs group, NRP-1 inhibitor plus RGE-modified NPs group, and unmodified NPs group. For the NRP-1 inhibitor plus RGE-modified NPs group, the cells were first cultured in the presence of NRP-1 inhibitor EG00229 (R&D Systems) at 100 μM. The three groups of cells were washed with phosphate buffered saline (PBS) and incubated with different concentrations of their corresponding NPs for 2 h.
To observe the cellular uptake of NPs qualitatively, the treated cells were washed three times with cold PBS, fixed with 4 % paraformaldehyde, stained with Hoechst33342 and observed using a confocal laser scanning microscope (CLSM, TCS SP5, Leica, Germany).
For quantitative analysis, the treated cells were washed with cold PBS, trypsinized and harvested by centrifugation at 1200 rpm for 5 min. The cells were resuspended in 200 μL PBS and filtered through a 40 mm nylon mesh to remove cell aggregates. The cell suspension was then analyzed by flow cytometry (Guava easyCyte, USA).
To observe the cellular uptake of NPs qualitatively, the treated cells were washed three times with cold PBS, fixed with 4 % paraformaldehyde, stained with Hoechst33342 and observed using a confocal laser scanning microscope (CLSM, TCS SP5, Leica, Germany).
For quantitative analysis, the treated cells were washed with cold PBS, trypsinized and harvested by centrifugation at 1200 rpm for 5 min. The cells were resuspended in 200 μL PBS and filtered through a 40 mm nylon mesh to remove cell aggregates. The cell suspension was then analyzed by flow cytometry (Guava easyCyte, USA).
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