Matrigel assay was performed as described previously.49 (link) Briefly, bEnd3 endothelial cells were plated in a six-well Matrigel plate (BD Biocoat Matrigel Matrix, 354432). The cells were seeded at a density of 105 cells per well in 2 mL medium. At 2 hours after seeding in Matrigel, the cells were treated with GSNO (100 μM), 2-ME (5 μM), or GSNO followed by 2-ME (30 minutes later). The cells were observed under a microscope for formation of tube-like structures at 3, 18, and 48 hours. Tubule length ratio, branch number per field, and aggregation area were analyzed using ImageJ software.
Biocoat matrigel matrix
Manufactured by BD
BD BioCoat Matrigel Matrix is a commercially available extracellular matrix product derived from a mouse sarcoma cell line. It provides a complex mixture of basement membrane proteins, including laminin, collagen IV, heparan sulfate proteoglycans, and entactin. This matrix supports the attachment, migration, differentiation, and survival of many cell types, including epithelial, endothelial, and certain stem cells.
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Lab products found in correlation
3 protocols using biocoat matrigel matrix
1
Matrigel Tubulogenesis Assay Protocol
2
Quantitative Matrigel Invasion Assay
3
Invasion Assay of Lung Cancer Cells
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For invasion assays, transiently transfected or stable knockdown of PPAT, PAICS and PKM2 in were used in A549 cells. BEAS-2B (benign transformed lung epithelial) cells were infected with PAICS-adenovirus. Seventy-two hours post-transfection and/or infection, cells were seeded onto BD BioCoat matrigel matrix (BD, CA) present in the insert of a 24-well culture plate. For A549, 10% serum containing RPMI medium was added to the lower chamber as a chemoattractant. For BEAS-2B, BEBM medium with supplementary growth factors was added as chemoattractant. After 36 h, the non-invading cells and matrigel matrix were gently removed with a cotton swab. Invasive cells located on the lower side of the chamber were stained with 0.2% crystal violet in methanol, air-dried and photographed. They were then enumerated microscopically using multiple representative areas. For colorimetric assays, the inserts were treated with 150 μl of 10% acetic acid and the absorbance measured at 560 nm [71 (link)]. Each experiment has been carried twice in triplicates per sample. Data shown is representative figure of one such experiment.
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