illation counter.
Currently, a sandwich-ELISA is also often used, which involves an antibody
coupled ELISA plate. The hGH protein binds to the antibody on the plate,
a digoxygenated antibody binds to hGH, and a secondary antibody coupled
to alkaline phosphatase is used for detection.
Green fluorescent protein
Green fluorescent protein (GFP), originally isolated from the jellyfish
Aequorea victoria (3), has the ability to absorb blue light and emit green
light. This unique protein can be expressed in mammalian cells and protein
expression can be visually monitored in living cells. Although the system
provides a convenient way to detect protein expression without a specific
assay, quantitative analysis is limited. This reporter gene system is best
suited for in situ detection of gene expression, such as localization studies
of fusion proteins within cells.
References
- Tang, M.X., Redemann, C.T., and Szoka, Jr F.C. (1996) In vitro gene
delivery by degraded polyamidoamine dendrimers. Bioconjugate Chem. 7,
703714.
- Freshney, R.I. (1994) Culture of Animal Cells, A Manual of Basic Technique,
Third Edition, Wiley-Liss.
- Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D. D., Seidman, J.
G., Smith, J. A., and Struhl, K., eds. (1991) Current Protocols in Molecular
Biology, Wiley Interscience, New York.
- Sambrook, J., Fritsch, E.F., and Maniatis, T., eds. (1989) Molecular
cloning a laboratory manual, 2nd Ed. Cold Spring Harbor Laboratory
Press.