TY - JOUR
T1 - Biochemical Fractionation of Membrane Receptors in the Nucleus
AU - Wang, Ying Nai
AU - Huo, Longfei
AU - Hsu, Jennifer L.
AU - Hung, Mien Chie
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2015.
PY - 2015
Y1 - 2015
N2 - Fractionation of cytoplasmic and nuclear proteins is a well-recognized biochemical technique to detect the intracellular distribution and expression level of proteins of interest. In the last decade, accumulating evidence shows that various types of cell surface receptors, such as receptor tyrosine kinases (RTKs), peptide hormone receptors, and cytokine receptors, are detected in the nuclei. Therefore, subcellular fractionation, including nonnuclear/nuclear extraction and the subsequent subnuclear fractionation without detectable crosscontamination during the process, is critical for studying membrane receptors that transit from the cell surface to the nucleus. Here, we utilize the epidermal growth factor receptor (EGFR) tyrosine kinase as an example of a comprehensive biochemical protocol for isolating membrane receptors in the nuclei of cancer cells.
AB - Fractionation of cytoplasmic and nuclear proteins is a well-recognized biochemical technique to detect the intracellular distribution and expression level of proteins of interest. In the last decade, accumulating evidence shows that various types of cell surface receptors, such as receptor tyrosine kinases (RTKs), peptide hormone receptors, and cytokine receptors, are detected in the nuclei. Therefore, subcellular fractionation, including nonnuclear/nuclear extraction and the subsequent subnuclear fractionation without detectable crosscontamination during the process, is critical for studying membrane receptors that transit from the cell surface to the nucleus. Here, we utilize the epidermal growth factor receptor (EGFR) tyrosine kinase as an example of a comprehensive biochemical protocol for isolating membrane receptors in the nuclei of cancer cells.
KW - Cell surface membrane receptor
KW - Epidermal growth factor receptor
KW - Immunoblotting
KW - Innernuclear membrane purifi cation
KW - Nuclear extraction
KW - Receptor tyrosine kinase
KW - Subnuclear fractionation
KW - Sucrose gradient centrifugation
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U2 - 10.1007/978-1-4939-1755-6_9
DO - 10.1007/978-1-4939-1755-6_9
M3 - Article
C2 - 25304351
AN - SCOPUS:84921788488
SN - 1064-3745
VL - 1234
SP - 99
EP - 112
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
ER -