The Design of a Quantitative Western Blot Experiment
Table 1
Major design parameters for protein-based assays. A solid experimental design that maps the procedure, controls, replicates, experimental conditions, and sampling handling guidelines sets the foundation for production of solid, quantifiable data.
Experiment procedure
Control groups
Replicates
Experiment conditions
Sample handling
Disease or treatment groups
Time course study (i.e., )
Biological (difft sample per well)
Growth conditions (media and time or OD)
Precise time to harvest cells or tissues
Target proteins implicated
Normal versus disease (i.e., normal)
Technical (same sample per well)
Days of embryonic development
Sample extraction method
Potential internal controls
Untreated versus drug treated (i.e., untreated)
Amount per mass of drug or compound
Preservation method and time
These first three steps define the following: experimental parameters and the goals and the samples based on literature or experimental data typically from broad microarray or proteomics experiments
