Native Proteins

Proteins

Native Proteins

Human

Biological activity/ Working concentration: Optimum concentration for human umbilical vein endothelial cells (HUVEC) range from 50-200 μg/ml

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

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)
PBS 1x
MeOH 2x 5 min
TCA 2x 10 min
H2O 1x
- 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)

inhibitors

optimal concentration with heparin (30 μg/ml) is about 12 μg/ml.

optimal concentration with heparin (50 μg/ml) is about 12 μg/ml.
Protocol: 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

Plasma

Spinal Cord

Brain

Liver

Placenta

Erythrocytes

Gizzard

Neutrophils

Stomach

Carcinoma

Cell culture

Egg white

Leukocytes

Pancreas

Pituitary

S. avidinii

Serum

Urine

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