Two Dimensional Substrates

Well-characterized protein-binding substrates, used extensively in other assays, have been transferred or "ported" for the use of microarray fabrication. Most of these surface substrates were used extensively in DNA microar-ray research and have been used with success with purified protein arrays. There are two principal ways that proteins bind to surfaces: (1) by the formation of covalent bonds with reactive groups on the surface of the slide, and (2) by noncovalent interactions with the slide substrate.

In slide surface chemistry, the use of aldehyde and epoxy coatings allows for covalent bonds between the proteins by virtue of the amine group on amino acids and the activated aldehyde of the epoxy group on the slide [1,2]. The primary amine group reacts with the aldehyde, or epoxy, groups on the slide surface. The electrons on the nitrogen attach to the carbon with a partial positive charge on the reactive group and form a nitrogen-carbon bond. In protein-aldehyde coupling, the attachment is stabilized by a dehydration reaction. Another option for two-dimensional substrates is the coat the slide surface with a molecule that is part of a high- affinity binding interaction. Binding-partner substrates are not as common but are still useful. There are examples of protein-binding surfaces that have been treated with avidin, strep-tavidin, glutathione, and monoclonal antitag antibodies. Unlike the aldehyde and epoxy examples, these are very specific and require that the appropriate binding partner be coupled to the protein(s) being spotted.

Project Management Made Easy

Project Management Made Easy

What you need to know about… Project Management Made Easy! Project management consists of more than just a large building project and can encompass small projects as well. No matter what the size of your project, you need to have some sort of project management. How you manage your project has everything to do with its outcome.

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