Nevertheless, it should be appreciated that antibody microarray-derived data is not as reliable as immunoblotting results, although it may be much more sensitive for target detection. Antibody microarray analyses are performed with non-denatured, native proteins. Consequently, there can be a very high rate of false-positive and false-negative results, i.e., the observed change in the expression or phosphorylation of an intended target for an antibody may not be reproduced by Western blotting. Antibodies can cross-react with off-target proteins. High density antibody microarrays rely on the immunocapture of proteins in crude cell lysates that have been labeled in vitro with a fluorescent dye. Since many proteins reside in complexes, it is possible that apparent changes in protein expression or phosphorylation can also reflect alterations in protein-protein interactions. False-negatives may result if the epitope recognized by an antibody on the microarray is not accessible when the target protein is complexed. Therefore, we strongly advise that extreme caution is exercised when considering KiNET-AM data. We highly recommend that any results from this database be confirmed by Western blotting before any further follow up is undertaken. This can be undertaken for clients with the Kinetworks™ Custom Immunoblotting services offered by Kinexus.
Both KiNET-IB and KiNET-AM have built in bioinformatics searching capabilities to retrieve data and generate custom data tables that are tailored to users’ specific cell signalling research questions. Both KiNET databases can be queried for the regulation of a target protein in hundreds of well-defined experimental model systems. Alternatively, a tissue, cell line or specific treatment can be interrogated for changes in the expression and phosphorylation of hundreds of possible proteins.
At this time, please cite the following reference for any data from the KiNET Data Bank that is reproduced elsewhere:
Pelech, S. et al. (2012) Kinetica Online – KiNET Data Bank (Kinexus Bioinformatics Corporation, Vancouver, BC, Canada)
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