Products

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

Human

E. coli

Human 293

Plasma

P.Pichia

Spinal Cord

Brain

Liver

Baculovirus

HEK293

Placenta

293F

Erythrocytes

Gizzard

Insect

Neutrophils

Stomach

CHO Stable

Carcinoma

Cell culture

Egg white

Expi293

Leukocytes

Mammalian

Pancreas

Pichia pastoris

Pituitary

S. avidinii

Serum

Synthetic

Urine

2

3