TY - JOUR
T1 - Protein microarrays to detect protein-protein interactions using red and green fluorescent proteins
AU - Kukar, Thomas
AU - Eckenrode, Sarah
AU - Gu, Yunrong
AU - Lian, Wei
AU - Megginson, Mike
AU - She, Jin Xiong
AU - Wu, Donghai
N1 - Funding Information:
1 This research is supported by NIH Grants DK58778 (J.X.S.) and GM58197 (D.H.W.). T.K. is supported by a University of Florida Alumni Fellowship.
PY - 2002/7/1
Y1 - 2002/7/1
N2 - Proteomics, the study of protein function on a global scale, will play an important role in furthering our understanding of gene functions, complex biological pathways, and discovery of novel drug targets. A number of techniques have been developed for proteomic studies to identify and analyze proteins, compare protein expression levels, and study protein-protein interactions. Recent developments have applied a DNA array-type approach to immobilize proteins on a surface for high-throughput analysis. Here we report the development and construction of protein chips using derivatized glass and nitrocellulose-coated slides and the employment of recombinant proteins fused with green and red fluorescent proteins for detection. Fluorescent signals were found to be proportional to the amount of arrayed proteins and could be readily detected with a conventional fluorescence slide scanner. This technique allows the investigation of protein-protein interactions without the need for additional labeling steps of probe proteins.
AB - Proteomics, the study of protein function on a global scale, will play an important role in furthering our understanding of gene functions, complex biological pathways, and discovery of novel drug targets. A number of techniques have been developed for proteomic studies to identify and analyze proteins, compare protein expression levels, and study protein-protein interactions. Recent developments have applied a DNA array-type approach to immobilize proteins on a surface for high-throughput analysis. Here we report the development and construction of protein chips using derivatized glass and nitrocellulose-coated slides and the employment of recombinant proteins fused with green and red fluorescent proteins for detection. Fluorescent signals were found to be proportional to the amount of arrayed proteins and could be readily detected with a conventional fluorescence slide scanner. This technique allows the investigation of protein-protein interactions without the need for additional labeling steps of probe proteins.
KW - Fluorescent proteins
KW - Green fluorescent protein
KW - Microarrays
KW - Protein interactions
KW - Red fluorescent protein
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U2 - 10.1006/abio.2002.5614
DO - 10.1006/abio.2002.5614
M3 - Article
C2 - 12069413
AN - SCOPUS:0036629196
SN - 0003-2697
VL - 306
SP - 50
EP - 54
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -