TY - CHAP
T1 - Protein arrays III
T2 - Reverse-phase protein arrays
AU - Yuan, Yulin
AU - Hong, Xia
AU - Lin, Zuan Tao
AU - Wang, Hongting
AU - Heon, Mikala
AU - Wu, Tianfu
PY - 2017
Y1 - 2017
N2 - The reverse-phase protein array (RPPA) is to use highly specific antibodies to interrogate pan or posttranslationally modified protein targets, such as phosphorylated proteins, particularly the proteins involved in cell signaling pathways. In this protocol we will cover the preparation of cell (or tissue) lysates, sample printing, antibody validation, antibody interrogation, signal amplification steps, imaging and data analysis. In this protocol, colorimetric catalyzed signal amplification (CSA) chemistry, fluorescence and near-infrared (NIR) based detection methods will be described.
AB - The reverse-phase protein array (RPPA) is to use highly specific antibodies to interrogate pan or posttranslationally modified protein targets, such as phosphorylated proteins, particularly the proteins involved in cell signaling pathways. In this protocol we will cover the preparation of cell (or tissue) lysates, sample printing, antibody validation, antibody interrogation, signal amplification steps, imaging and data analysis. In this protocol, colorimetric catalyzed signal amplification (CSA) chemistry, fluorescence and near-infrared (NIR) based detection methods will be described.
KW - Antibody validation
KW - Cell signaling pathways
KW - Near-infrared (NIR) fluorescence
KW - Reverse-phase protein array
UR - http://www.scopus.com/inward/record.url?scp=85030751583&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85030751583&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-7231-9_21
DO - 10.1007/978-1-4939-7231-9_21
M3 - Chapter
C2 - 28986799
AN - SCOPUS:85030751583
VL - 1654
T3 - Methods in Molecular Biology
SP - 279
EP - 289
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -