Weaknesses
The biggest limitation of RPMA, as is the case for all immunoassays, is its dependence on antibodies for detection of proteins. Currently there is a limited but rapidly growing number of signaling proteins for which antibodies exist that give an analyzable signal. In addition, finding the appropriate antibody could require extensive screening of many antibodies by western blotting prior to beginning RPMA analysis. To overcome this issue, two open resource databases have been created to display western blot results for antibodies that have good binding specificity within the expected range (http://discover.nci.nih.gov/abminer and http://mtt.uscancer.org). Furthermore, RPMAs, unlike western blots, do not resolve protein fractions by molecular weight. Thus, it is critical that upfront antibody validation be performed.
Read more about this topic: Reverse Phase Protein Lysate Microarray
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