Product Description for Endothelial Cell Growth Factor (ECGFpro1)
Properties for Endothelial Cell Growth Factor (ECGFpro1)
|Product Category||Proteins & Growth Factors|
|PDF datasheet||View Datasheet|
|Manufacturer||Acris Antibodies GmbH|
|Background||Endothelial cells from human umbilical vein (HUVEC) can be established as primary cultures by traditional methods. The serial propagation of these cells has proved to be difficult. The long-term propagation of these cells in vitro can be achieved with an extract prepared from bovine brain. The introduction of a fibronectin or collagen matrix to the cell culture system allows to cultivate endothelial cells at clonal densities. With ECGF, the FCS requirement can be reduced. Heparin potentiates the mitogenic activity of crude preparations of ECGF. ECGF has also been reported to eliminate the need for feeder cells in the clonal growth of hybridomas and other cell types.|
|Protocols||Thymidine Incorporation with HUVEC
- plate cells with a density at 5-7 x 10e3 cells/well in growth medium (EGM)
- incubate cells over night [if urgent, plate cells in the morning, change growth medium against basal medium (EBM) in the early afternoon]
- change EGF against EBM (for HUVEC: EBM +1-2% FCS)
- incubate 24 h
- change medium again and add factors (growth factors, inhibitors, etc.)
- incubate for 18 h
- add 10 µl 3H-Thymidine solution [0.025 mCi/ml] per well (=0.25 µCi)
- incubate another 6h at 37°C
- Washing steps: (250 µl/well)
MeOH 2x 5 min
TCA 2x 10 min
- lyse cells in 250 µl 0.3M NaOH per well
- transfer 2.5 ml ECO Plus into the appropriate scintillation vials
- transfer cell lysats into the scintillation vials
- count by liquid scintillation (ß-counter; Beckmann Instruments)
|Storage||Store lyophilized at 2-8°C for 6 months or at -20°C long term.
After reconstitution store the antibody undiluted at 2-8°C for one month
or (in aliquots) at -20°C long term.
Avoid repeated freezing and thawing.
Shelf life: one year from despatch.
|Source||Bovine brain (BSE-free tested!)|