TY - JOUR
T1 - Methods for immunoblot detection and electron microscopic localization of septin subunits in mammalian nervous systems
AU - Parajuli, L. K.
AU - Ageta-Ishihara, N.
AU - Ageta, H.
AU - Fukazawa, Y.
AU - Kinoshita, M.
N1 - Publisher Copyright:
© 2016 Elsevier Inc.
Copyright:
Copyright 2016 Elsevier B.V., All rights reserved.
PY - 2016
Y1 - 2016
N2 - The minimal functional units of the mammalian septin system are diverse heterooligomers of SEPT1-14 subunits, which are most abundantly and differentially expressed in postmitotic neurons and glia. The subunit compositions of such heterooligomers are thought to differentiate their affinity for other proteins and lipids, and subcellular localization. Thus, high-precision quantification and mapping of each subunit is necessary to understand their subcellular functions and physiological roles. However, systematic information on the localization of individual septin subunits in the mammalian nervous system is limited. Here, we present our experimental workflows for the study of septin expression and localization in the rodent brain by immunoblot and serial section immunoelectron microscopy. Our protocols, based on standard methods, have been rigorously optimized and simplified for universality and reproducibility to aid non-experts in the field.
AB - The minimal functional units of the mammalian septin system are diverse heterooligomers of SEPT1-14 subunits, which are most abundantly and differentially expressed in postmitotic neurons and glia. The subunit compositions of such heterooligomers are thought to differentiate their affinity for other proteins and lipids, and subcellular localization. Thus, high-precision quantification and mapping of each subunit is necessary to understand their subcellular functions and physiological roles. However, systematic information on the localization of individual septin subunits in the mammalian nervous system is limited. Here, we present our experimental workflows for the study of septin expression and localization in the rodent brain by immunoblot and serial section immunoelectron microscopy. Our protocols, based on standard methods, have been rigorously optimized and simplified for universality and reproducibility to aid non-experts in the field.
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U2 - 10.1016/bs.mcb.2016.04.021
DO - 10.1016/bs.mcb.2016.04.021
M3 - Article
C2 - 27473915
AN - SCOPUS:84999622413
VL - 136
SP - 285
EP - 294
JO - Methods in Cell Biology
JF - Methods in Cell Biology
SN - 0091-679X
ER -