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Proteomic studies of human and mouse tubulin typically result in the analysis of a large number of proteins, many of which are specific isoforms. Their identification by mass spectrometry (MS) depends on efficient sample preparation and separation. Proteomic studies of tubulin have taken advantage of its interaction with F-actin and peptide microtubule stabilizing agents, leading to the identification of binding partners. One approach is based on chemical crosslinking of purified tubulin with glutaraldehyde. The resulting cross-linked polymers are purified by affinity chromatography using tubulin antibodies as a probe. Peptides resulting from the digestion of the cross-linked polymers are analysed by MS/MS fragmentation and results are matched to a database. The approach is powerful, but it also has several drawbacks. The use of antibodies to isolate crosslinked polymers is a major source of errors in the identification process, which depend on the specificity of the antibodies used and the type of proteins targeted. This is illustrated here by the analysis of the tubulin-F-actin microtubule cross-linking. We also review briefly two alternative approaches to affinity purification of tubulin complexes: chemical cross-linking and methods based on reversible chemical modification of the beta-tubulin thiols. Finally, we describe in more detail the use of LysC and Trypsin in the preparation of samples for MS/MS analysis.This is a great site for real-time traffic, news, and weather data for the greater Toronto area.
Managing plugin for each timezone adds a little overhead to each timezone page, but the traffic data on this site is pretty much all related to traffic around Toronto, so adding the timezone data to this site makes sense.
This site has a lot of great features for seeing at a glance where your bike is in real-time, wherever you are. There are some limitations to it, which are outlined on the about page.
This is a great site for counting bicycle traffic and measuring cycling levels around Toronto. It also has an Ontario-wide location-based bicycle route planner.
One of the most popular iPhone apps of the moment. Allows you to search bike parking racks
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