Fabrication of protein function microarrays for systems-oriented proteomic analysis.
Protein microarrays have many potential applications in high-throughput analysis of protein function. However, simple, reproducible, and robust methods for array fabrication are required. Here we discuss the background to different routes to array fabrication and describe in detail one approach in which the purification and immobilization procedures are combined into a single step, dramatically simplifying the array fabrication process. We illustrate this approach by reference to the creation of an array of p53 variants, and discuss methods for assay and data analysis on such arrays.
Blackburn
Jonathan M
JM
Department of Biotechnology, University of the Western Cape, Cape Town, South Africa.
Hart
Darren J
DJ
D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D010957:Plasmids; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D011485:Protein Binding; D011487:Protein Conformation; D040901:Proteomics / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
D008024:Ligands; D011993:Recombinant Fusion Proteins; D016159:Tumor Suppressor Protein p53