Data Bank

KiNET Data Bank - Repositories of Signal Transduction Protein Expression and Phosphorylation Data from Kinexus Proteomics Services

Kinexus Bioinformatics Corporation has provided Kinetworks™ Multi-immunoblotting and Kinex™ Antibody Microarray services with over 850 highly-tested antibodies to quantify signal transduction protein expression and phosphorylation since the company's incorporation in 1999. All of the protein measurements deposited in the KiNET Data Bank were generated with the top 22% of over 4500 commercial antibodies that were independently tested and validated by Kinexus. Over 15,000 tissue and cell specimens from thousands of human and experimental animal model systems have been submitted to Kinexus from over 1650 clients for proteomics analyses with these antibodies. With the permission of these clients, much of this unique data is freely available from the KiNET Data Bank to other academic and industrial biomedical scientists to benefit their own research programs. Over 95% of the data in the KiNET Data Bank is unpublished and not retrievable elsewhere.

The KiNET – Immunoblotting DataBase (KiNET-IB) features over 200,000 measurements of the expression and phosphorylation states of hundreds of cell signalling target proteins from over 6000 full Kinetworks™ multi-immunoblots blots performed with over 4000 control and treated tissue/cell samples. Since over the last 14 years, all of the KiNET–IB data was produced with the same reagents, methodology and equipment by the scientists and technicians at Kinexus, the results are highly comparable. Moreover, as each measurement is based on both the immunoreactivity and molecular mass of the target protein, this data is amongst the most reliable available today for specific protein levels and phosphorylation in a large number of diverse biological specimens.

The KiNET-Antibody Microarray DataBase (KiNET-AM) holds the quantitative results from nearly 2000 Kinex™ Antibody Microarray analyses with over 1.5 million measurements of over 700 different signalling proteins and phospho-sites. Each measurement is the average of duplicate determinations. Only single-dye, non-competitive methodology was used to generate the data in KiNET-AM (i.e., all protein samples were labeled with the same dye and incubated separately from each other on microarrays). This greatly reduces the occurrence of misleading results.
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)

Click here to open KiNET-IB

Click here to open KiNET-AM