Education
Application of Reverse Phase Protein Arrays
Cell Lines
- Characterize cellular signaling networks under different culture conditions
- Determine drug selectivity
- Identify therapeutic targets
- Define regulatory mechanisms in signaling networks, including forward and feedback loops and cross-talk
- Ability to analyze up to 1000 conditions concurrently allows the detailed concentration and time courses required for systems biology approaches
Patient Samples
- Classify patient tumors
- Correlate DNA, RNA and protein
- Determine prognosis
- Predict responses to targeted therapies
- Pharmacodynamics and biologically relevant dose
- Determine appropriate handling procedures for clinical samples (based on antigen stability analysis)
In Development
- Laser-aided microdissection validation for RPPA
- Paraffin embedded tumors validation for RPPA
Antibody Validation
- A single or dominant band on western blotting is required
- Dynamic range and specificity is determined using peptides, phosphopeptides, growth factors, inhibitors, RNAi or cells with wide levels of expression
- Three hundred thirty cell lines under multiple conditions on a single array for antibody validation
- A Pearson correlation coefficient between RPPA and western blotting of greater than 0.7 is required
- Each new antibody batch is validated
- Approximately 500 antibodies under validation process
Please refer to the list of RPPA-validated antibodies.
Validated Antibodies
Our validated antibody list (xls) was last updated December 2009.