Proteome Analysis
In Vitro translation - can synthesize proteins from mRNA
Wheat Germ
Plant extract that contains ribosomes, tRNA,
Reticulocyte
Using nuclease, most mRNA can be removed. Also has a energy regenerating system.
Minicells
Anucleated cells that contain all the machinery to make proteins from mRNA taken up by the cells.
Protein-Protein interactions
-Yeast two-hybrid system.
The DNA binding domains and Activation domains are separate in transcription factors.
The separate domains can be cloned onto proteins that are to be selected for protein-protein interaction.
-Phage display
Protein Analysis
-Size => SDS Polyacrylamide Gel Electrophoresis (SDS-PAGE)
-Charge => Isoelectric Focusing Gel Electrophoresis (IEF-PAGE)
-2D Gels combine IEF-PAGE and SDS-PAGE
-Separation technology =>
Columns
Electrophoresis (2D gels)
HPLC
-Native state, subunits => Native Gel Electrophoresis, Analytical Centrifugation
-Amino Acid Analysis => Degradation, separation on Columns
-Protein Sequence => Sequential degredation and HPLC
-Protein Identification =>
Quantifying and selection of protein
An antigen is a molecule that can elicit an immune response and an epitope is the specific portion of the molecule that the immune response is eleicited agains.
Antibodies can be tagged with enzymes such as alkaline phosphatase and horseradish peroxidase.
-Elisa
-Immunoprecipitation
-Western Blots
