Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 327-333 |
| Number of pages | 7 |
| Journal | Histochemistry and Cell Biology |
| Volume | 120 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Oct 2003 |
| Externally published | Yes |
Fingerprint
Keywords
- Avidin-FITC
- Avidin-interacting proteins
- Avidin-peroxidase
- Cultured cells
- Mitochondria
ASJC Scopus subject areas
- Cell Biology
- Instrumentation
Cite this
Transblot and cytochemical identification of avidin-interacting proteins in mitochondria of cultured cells. / Sasaki, Eiko; Okamoto, Yoshifumi; Yoshida, Kaya; Okamura, Hirohiko; Shimizu, Katsuhide; Nasu, Fumio; Morimoto, Hiroyuki; Haneji, Tatsuji.
In: Histochemistry and Cell Biology, Vol. 120, No. 4, 10.2003, p. 327-333.Research output: Contribution to journal › Article
}
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
Y1 - 2003/10
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
UR - http://www.scopus.com/inward/record.url?scp=0242361073&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0242361073&partnerID=8YFLogxK
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 - Histochemistry and Cell Biology
JF - Histochemistry and Cell Biology
SN - 0948-6143
IS - 4
ER -