Matrigel plug angiogenesis assay (Corning, NY, USA) was used to evaluate the in vivo anti-angiogenic effect of miRNA-124. The matrigel stock solution was thawed overnight at 4°C. A gel solution was prepared using a Matrigel stock solution and serum-free DMEM, and the solution was placed in a 96-well plate and then allowed to incubate for 2 hrs to cure. The cultured cells were collected and digested to prepare a single cell suspension under aseptic conditions, and the cell suspension was adjusted to a density of 1×105/mL. The cells were seeded in 96-well plates at 100 μL per well. The plates were incubated for 6–8 hrs in an incubator (5% CO2, 37°C), and the cells were visualized using an inverted microscope (Thermo Fisher Scientific) and photographed.
Matrigel plug angiogenesis assay
Manufactured by Corning
Sourced in United States
Matrigel plug angiogenesis assay is a laboratory technique used to study the process of angiogenesis, which is the formation of new blood vessels. It involves the use of a gelatinous protein mixture, Matrigel, which provides a three-dimensional matrix for the study of cell behavior, including the migration and differentiation of endothelial cells, which are essential for the formation of new blood vessels.
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4 protocols using matrigel plug angiogenesis assay
1
In Vivo Angiogenesis Assessment Using Matrigel Plug Assay
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In Vivo Angiogenesis Assay using Matrigel
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Matrigel plug angiogenesis assay (Corning, NY, USA) was used to determine in vivo angiogenic rate. Matrigel stock solution was thawed overnight at 4 °C. A gel solution was prepared using a Matrigel stock solution and serum-free DMEM, and the solution was placed in a 96-well plate and then allowed to be incubated for 2 h to cure. The cultured cells were collected and digested to prepare a single cell suspension under aseptic conditions, and the cell suspension was adjusted to a density of 1×105/mL. The cells were seeded in 96-well plates at 100 μL per well. The plates were incubated for 6–8 h in an incubator (5% CO2, 37 °C) and the cells were visualized using an inverted microscope (Thermo Fisher Scientific) and then photographed.
3
Matrigel Plug Angiogenesis Assay for ELF5 Anti-angiogenic Evaluation
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Matrigel plug angiogenesis assay (Corning, NY, USA) was used to evaluate the in vitro anti-angiogenic effect of ELF5. The Matrigel stock solution was thawed overnight at 4°C. A gel solution was prepared using a Matrigel stock solution and serum-free RPMI-1640 medium, and the solution was placed in a 96-well plate and then allowed to incubate for 2 h to cure. The cultured SKOV-3 cells were collected and digested to prepare a single-cell suspension under aseptic conditions, and the cell suspension was adjusted to a density of 1×105/ml. The cells were seeded in 96-well plates at 100 μl per well. The plates were incubated for 6–8 h in an incubator (5% CO2, 37°C), and the cells were visualized using an inverted microscope (Thermo Fisher Scientific) and photographed.
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Matrigel Plug Angiogenesis Assay for Anti-Angiogenic Evaluation
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We used Matrigel plug angiogenesis assay (Corning, NY, USA) to evaluate the in vivo anti-angiogenic effect of TTAC-0001. It is based on the fact that VEGF secreted from Matrigel tumor cells exerts pro-angiogenic effects on the surrounding tissue, thus promoting neovascular development that is visualized in the Matrigel. If a drug inhibits the effect of VEGF by inhibiting either VEGF or VEGFR, then the decreased levels of neovascular development in the Matrigel can be evaluated.
Matrigel (0.5 mL) was premixed with 5 × 106 MDA-MB-231 cells and then engrafted into the right mammary fat pad of Balb/c nude mice (n = 5). A single 2 or 10 mg/kg treatment of TTAC-0001 or saline was administered to the respective groups by intravenous injection after implantation. After 10 days, Matrigel plugs were removed and frozen for immunofluorescence analysis. To measure the hemoglobin (Hb) content, excised plugs (n = 5 plugs/group) were cut into small pieces and placed in 500 uL of cold, distilled water at 4°C overnight to liquefy the Matrigel. Specimens were centrifuged at 1500 rpm for 20 mins, and the supernatant was collected. Hb content was quantified using a Hb assay kit (Sigma-Aldrich) and spectrophotometry.
Matrigel (0.5 mL) was premixed with 5 × 106 MDA-MB-231 cells and then engrafted into the right mammary fat pad of Balb/c nude mice (n = 5). A single 2 or 10 mg/kg treatment of TTAC-0001 or saline was administered to the respective groups by intravenous injection after implantation. After 10 days, Matrigel plugs were removed and frozen for immunofluorescence analysis. To measure the hemoglobin (Hb) content, excised plugs (n = 5 plugs/group) were cut into small pieces and placed in 500 uL of cold, distilled water at 4°C overnight to liquefy the Matrigel. Specimens were centrifuged at 1500 rpm for 20 mins, and the supernatant was collected. Hb content was quantified using a Hb assay kit (Sigma-Aldrich) and spectrophotometry.
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