RPP1C Antibody

Role of Antibodies in RPPA Technology

RPPA employs antibodies to quantify protein levels in biological samples spotted on nitrocellulose arrays. Each antibody’s specificity directly impacts data reliability . Key characteristics include:

• Target specificity: Antibodies must recognize a single epitope (e.g., phosphorylated residues, isoforms) .

• Validation: MD Anderson Cancer Center’s RPPA Core Facility categorizes antibodies as Valid, Use with Caution, or Under Evaluation based on Western blot correlation (>0.7 Pearson) and single-band confirmation .

• Applications: Used in cancer research to profile signaling pathways (e.g., EGFR, mTOR) and therapeutic responses .

Antibody Validation and Reliability

RPPA antibody performance correlates with mRNA-protein consistency:

Lower-reliability antibodies reduce correlation accuracy due to cross-reactivity or nonspecific binding .

Multi-Platform Concordance of RPPA Antibodies

Studies comparing RPPA platforms show:

• Inter-antibody consistency: Antibodies targeting the same antigen (e.g., phosphorylated Erk1/2) cluster closely (Spearman ρ = 0.22–0.75) .

• Cross-platform reproducibility: Integrative analyses of breast cancer cell lines treated with kinase inhibitors revealed concordant pathway activation profiles across different antibodies and platforms .

Challenges and Recommendations

• Antibody selection: Prioritize antibodies with Valid status and peer-reviewed applications .

• Data interpretation: Pair RPPA findings with orthogonal methods (e.g., mass spectrometry) to confirm targets .

• Limitations: "Tissue-reactive" antibodies may produce false signals in human tissue samples .