RESEARCH TOOLS & PROTOCOLS
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Culture Primary Endothelial Cells M1168 is tested and optimized with mouse endothelial cell growth and proliferation. Use aseptic technique to prevent microbial contamination.
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Freezing or Recovering Cells Cryo-preserved cells must be stored in liquid nitrogen or seeded immediately upon arrival. Coat sterile culture dishes or flasks with Gelatin-Based Coating Solution (Cell Biologics, Catalog No. 6950) for 2 min and then aspirate the excess solution before seeding cells.
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Handling Live Cultured Cells Keep a flask with 20 ml existing medium in 37°C CO2 incubator for 1 hour before replacing the medium. Either split the 90% confluent cells from a T25 flask to a T75 flask after 1 hour or let the cells grow in the T25 flask for 12-24 hours before split the cells to a T75 flask. Cells should be checked daily under a microscopy to verify appropriate cell morphology.
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Protocol for Flow Cytometry, Uptake of Dil-Ac-LDL and Tube Formation Assay and cell staning (Endothelial Cells) For best results, please carefully review the detailed protocol to use an unconjugated primary antibody. *Incubate endothelial cells with Dil-Ac-LDL for 4 h at 37°C. *Thaw reduced growth factor matrigel (BD: 354234) by submerging the bottle on ice and store the matrigel at 4 °C overnight.
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FAQ about Custom Endothelial Cell Isolation Primary endothelial cells produced from Cell Biologics display typical "cobblestone" or "spindle-shiped" morphology under light microscopy and show VE-cadherin (CD144) staining at cell-cell junction or Von Willebrand Factor staining in plasma (see our website). These cells also express VE-cadherin and CD31 as demonstrated by FACS analysis.
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