Cell pellets were collected after trypsinization, washed with 1× PBS, and lysed with RIPA buffer (Thermo Scientific, 89900) containing protease inhibitors. Twenty to fifty micrograms of total cell lysates were resolved on SDS-PAGE gels and transferred onto PVDF membranes. Blots were probed with the following antibodies at the indicated dilutions; OxPhos antibody cocktail (Abcam, ab110411; 1:500 dilution), anti-Actin (Sigma, A5316; 1:1000), anti-NDUFB6 (Abcam, ab110244; 1:1000), anti-MTCO2 (Abcam, ab110258; 1:1000), anti-GAPDH (Millipore Sigma, ABS16; 1:1000), anti-VDAC1 (Abcam, ab14734; 1:1000), and anti-TOP1MT-3 (Developmental Studies Hybridoma Bank, CPTC-TOP1MT-3; 1:200). Appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies were used at 1:3000 as follows: goat anti rabbit IgG, HRP-linked Antibody (Cell Signaling Technology, 7074S), or goat anti-mouse IgG-HRP (Santa Cruz Biotechnology, sc-2055). Blots were incubated with Clarity ECL substrate (Biorad, 1705061) and imaged using an Amersham Imager AI600.
Anti top1mt 3
Manufactured by Developmental Studies Hybridoma Bank
Anti-TOP1MT-3 is a monoclonal antibody produced by the Developmental Studies Hybridoma Bank. It is designed to detect the topoisomerase I, mitochondrial (TOP1MT) protein. The antibody can be used for various research applications that require the identification and analysis of the TOP1MT protein.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (4 mentions)
Oxphos antibody cocktail, by Abcam (3 mentions)
Goat anti mouse igg hrp, by Santa Cruz Biotechnology (3 mentions)
Anti actin, by Merck Group (3 mentions)
Ai600 imager, by Cytiva (3 mentions)
Anti mtco2, by Abcam (3 mentions)
Clarity ecl substrate, by Bio-Rad (3 mentions)
Anti gapdh, by Merck Group (3 mentions)
Goat anti rabbit igg hrp linked antibody, by Cell Signaling Technology (2 mentions)
Anti vdac1, by Abcam (1 mentions)
Hrp linked antibody, by Cell Signaling Technology (1 mentions)
Cptc top1mt 3, by Developmental Studies Hybridoma Bank (1 mentions)
Anti ndufb6, by Abcam (1 mentions)
3 protocols using anti top1mt 3
1
Immunoblot Analysis of Mitochondrial Proteins
2
Western Blot Analysis of Mitochondrial Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
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RIPA buffer (Thermo Scientific™, 89 900) containing protease inhibitors was used to lyse collected cell pellets after washing with 1× PBS. SDS-PAGE gels were used to resolve 50 μg of total cell lysates from different cell lines then polyvinylidene fluoride (PVDF) membranes were used for the transferred blot. Blots were probed with several antibodies at the indicated dilutions; OxPhos antibody cocktail (Abcam, ab110411; 1:500 dilution), anti-Actin (Sigma, A5316; 1:1000), anti-MTCO2 (Abcam, ab110258; 1:1000), anti-GAPDH (Millipore Sigma, ABS16; 1:1000), anti-TOP1MT-3 (Developmental Studies Hybridoma Bank, CPTC-TOP1MT-3; 1:200). Appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies were used at 1:3000 as follows: goat anti rabbit IgG, HRP linked Antibody (Cell Signaling Technology, 7074S) or goat anti-mouse IgG-HRP (Santa Cruz Biotechnology, sc-2055). The Clarity ECL substrate (Biorad, 1 705 061) was used to expose the horseradish enzyme attached to each antibody using an Amersham Imager AI600.
3
Western Blot Analysis of Mitochondrial Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RIPA buffer (Thermo Scientific™, 89900) containing protease inhibitors was used to lyse collected cell pellets after washing with 1× phosphate buffered saline (PBS). SDS-PAGE gels were used to resolve 50 g of total cell lysates from different cell lines then PVDF membranes were used for the transferred blot. Blots were probed with several antibodies at the indicated dilutions; OxPhos antibody cocktail (Abcam, ab110411; 1:500 dilution), anti-Actin (Sigma, A5316; 1:1000), anti-MTCO2 (Abcam, ab110258; 1:1000), anti-GAPDH (Millipore Sigma, ABS16; 1:1000), anti-TOP1MT-3 (Developmental Studies Hybridoma Bank, CPTC-TOP1MT-3; 1:200). Appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies were used at 1:3000 as follows: goat anti rabbit IgG, HRP linked Antibody (Cell Signaling Technology, 7074S), or goat anti-mouse IgG-HRP (Santa Cruz Biotechnology, sc-2055). The Clarity ECL substrate (Biorad, 1705061) was used to expose the horseradish enzyme attached to each antibody using an Amersham Imager AI600.
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