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Lecture 2
Two types of cell culture: Cell Suspension & Monolayer Cell Culture
MOLECULAR TECHNIQUES
Responses to toxicants involves gene expression analyzed by
microarray
Studies of polymorphisms in xenobiotic-metabolizing and other
enzymes
Analysis of chemically induced mutations
Development of knockout animals lacking a particular gene
Transgenic animals with an additional transgene
Toxic effects on transcription, mRNA processing and translation
and protein synthesis
MOLECULAR TECHNIQUES
1. Molecular Cloning (Insertion of a DNA segment into a suitable
vector)
2. cDNA and Genomic libraries (used in many screening
procedures and many transgenic proteins)
3.
IMMUNOCHEMICAL TECHNIQUES
Immunolocalization:
is a technique for identifying the presence of a
protein within the cell, its relative abundance
and its subcellular localization
Immunoaffinity Purification
Involves the use of antibodies, bound to
an insoluble matrix, for chromatography.
Immunoprecipitation
immunoaffinity
means
to
is
purification
remove
variant
and
protein
of
is
from
Western Blotting:
is a widely used technique in which antibodies
are used to detect proteins following
electrophoresis