TY - JOUR
T1 - Transblot and cytochemical identification of avidin-interacting proteins in mitochondria of cultured cells
AU - Sasaki, Eiko
AU - Okamoto, Yoshifumi
AU - Yoshida, Kaya
AU - Okamura, Hirohiko
AU - Shimizu, Katsuhide
AU - Nasu, Fumio
AU - Morimoto, Hiroyuki
AU - Haneji, Tatsuji
PY - 2003/10/1
Y1 - 2003/10/1
N2 - Cell lysates prepared from 3T3-L1 cells were analyzed by western blotting using the avidin-biotin complex system and anti-Bax antibody. The antibody interacted with bands of proteins with estimated molecular weights of 120, 74, 72, and 25 kDa. However, only the 25-kDa band was detected with the anti-Bax antibody when the direct immunoblotting method was used. Peroxidase-conjugated avidin interacted with the 120-, 74-, and 72-kDa bands. This interaction was not limited to 3T3-L1 cells, because peroxidase-avidin also interacted with these three proteins in MC3T3-E1, YROS, Saos-2, MG63, SCCKN, and SCCTF cells although the staining intensity was different in each cell type. Avidin-peroxidase also interacted with these three proteins in the mitochondria-containing fractions prepared from 3T3-L1 cells. FITC-streptavidin was also localized in mitochondria in the cultured cells. The localization of avidin/ streptavidin-interacting proteins in mitochondria was confirmed by using double staining with FITC-streptavidin and Mito-tracker.
AB - Cell lysates prepared from 3T3-L1 cells were analyzed by western blotting using the avidin-biotin complex system and anti-Bax antibody. The antibody interacted with bands of proteins with estimated molecular weights of 120, 74, 72, and 25 kDa. However, only the 25-kDa band was detected with the anti-Bax antibody when the direct immunoblotting method was used. Peroxidase-conjugated avidin interacted with the 120-, 74-, and 72-kDa bands. This interaction was not limited to 3T3-L1 cells, because peroxidase-avidin also interacted with these three proteins in MC3T3-E1, YROS, Saos-2, MG63, SCCKN, and SCCTF cells although the staining intensity was different in each cell type. Avidin-peroxidase also interacted with these three proteins in the mitochondria-containing fractions prepared from 3T3-L1 cells. FITC-streptavidin was also localized in mitochondria in the cultured cells. The localization of avidin/ streptavidin-interacting proteins in mitochondria was confirmed by using double staining with FITC-streptavidin and Mito-tracker.
KW - Avidin-FITC
KW - Avidin-interacting proteins
KW - Avidin-peroxidase
KW - Cultured cells
KW - Mitochondria
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U2 - 10.1007/s00418-003-0572-x
DO - 10.1007/s00418-003-0572-x
M3 - Article
C2 - 14574588
AN - SCOPUS:0242361073
VL - 120
SP - 327
EP - 333
JO - Zeitschrift für Zellforschung und Mikroskopische Anatomie. Abteilung Histochemie
JF - Zeitschrift für Zellforschung und Mikroskopische Anatomie. Abteilung Histochemie
SN - 0948-6143
IS - 4
ER -