VEGF Secretion Quantification in Astrocytes

The level of VEGF secreted was assessed using a Mouse VEGF Immunoassay kit (R&D Systems) as we previously described [18 (link)]. Astroglial cells (6 x 10⁵) were plated on a 60 mm tissue culture dish. Once the cells had reached ~90% confluence they were rinsed with serum-free DMEM and then grown in serum-free DMEM for 2 days. The conditioned medium was then clarified. The VEGF Immunoassay was performed in triplicate for the samples and normalized to the number of cells. A standard curve was generated to assess the sample VEGF levels. The assay was repeated twice with similar results.

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Falero-Perez J., Sheibani N, & Sorenson C.M. (2020). Bim expression modulates the pro-inflammatory phenotype of retinal astroglial cells. PLoS ONE, 15(5), e0232779.

Publication 2020

Mouse VEGF Immunoassay kit

Assay Astroglial cells Cells Conditioned medium Dish Immunoassay Mouse Serum Tissue Vegf

Corresponding Organization : Smith-Kettlewell Eye Research Institute

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Level of VEGF secreted

control variables

Astroglial cells (6 x 10^5) plated on a 60 mm tissue culture dish
Cells reached ~90% confluence before being rinsed with serum-free DMEM and grown in serum-free DMEM for 2 days
The conditioned medium was then clarified

controls

A standard curve was generated to assess the sample VEGF levels
The assay was repeated twice with similar results

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