Microarrays membrane proteins

Fig. 2 Common types of microarrays DNA microarray (a), carbohydrate microarray (b), antibody microarray (c), functional protein microarray (d), biomembrane and membrane protein microarray (e), and cellular microarray (f)...

Fang Y, Frutos AG, Lahiri J (2002) Membrane protein microarrays. J Am Chem Soc 124 2394-2395... 

In this chapter, we will survey the kinds of solid supports (substrates) and surface chemistries currently used in the creation of nucleic acid and protein microarrays. Which are the best supports and methods of attachment for nucleic acids or proteins Does it make sense to use the same attachment chemistry or substrate format for these biomolecules In order to begin to understand these kinds of questions, it is important to briefly review how such biomolecules were attached in the past to other solid supports such as affinity chromatography media, membranes, and enzyme-linked immxm-osorbent assay (ELISA) microtiter plates. However, the microarray substrate does not share certain unique properties and metrics with its predecessors. Principal among these are printing, spot morphology, and image analysis they are the subjects of subsequent chapters. 

And, how is it that membranes cast upon glass substrates are now used to prepare protein microarrays We will address these questions in good time. 

Membranes such as NC supported on glass may be more applicable for protein microarrays than glass substrates. Supported charged nylon membranes for microarrays are currently entering the marketplace as well. The essential ingredient for protein is water. Protein hydration reduces the likelihood for surface denaturation. Hydrophilic membranes allow proteins to... 

Table 6.1 Properties of Commercial Membranes Useful as Protein Microarray Supports... 

In order to assess the quality of the p53 protein microarrays printed on SAM2 membranes, two different assays are carried out on replica arrays. The antibodybinding assay comprises binding a Cy3-labeled anti-His antibody to the arrayed p53 proteins. This assay is independent of protein activity but provides a measurement of the relative amounts of protein immobilized in each spot. 

In principle, any microarray scanner can be used to read the protein microarrays, since the differences lie mainly in the sensitivity. However, models that read in a transmission mode might not work with membrane-based arrays. 

Protein assays using ordered arrays emerged in the late 90s [13]. Recently Lahiri et al. introduced a technique which enables the fabrication of microarrays for membrane proteins [14]. Printed lipid microspots on y-aminopropylsilane slides have high mechanical stability, independent of whether the lipid is in the gel-or fluid-phase. They immobilized G-protein coupled receptors and studied their interaction with a set of inhibitors. 

While glass has proven to be the most popular conventional platform for protein microarrays, a number of other physical substrates have also been evaluated. These have included gold, silica, and polyvinylidene fluoride and nitrocellulose membranes. Nitrocellulose has been employed in array development as it can retain large quantities of antibody, is inexpensive, and disposable. Flowever, it is not suitable for kinetic studies due to the slow diffusion of proteins through its pores. It is also not compatible... 

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