Cells were washed with phosphate-buffered saline (PBS) and homogenized in cell lysis buffer (150 mM NaCl, 1% NP40, 10% DOC, 10% SDS, 50 mM Tris, pH 7.4). Determination of protein concentration was performed using the BCA protein assay kit (Pierce). Samples were mixed with standard Laemmli loading buffer and 30 μg protein loaded on mini-Protean TGX precast gel (Bio-Rad). After SDS-PAGE, proteins were transferred to Immunoblot PVDF membranes (BioRad), the membrane blocked with 3% BSA, and further incubated with rabbit monoclonal anti-DIMT1 antibody (1:1000 dilution) and Rabbit polyclonal anti-OXPHOS (1:250 dilution antibody; both from Abcam), anti-NBR1 and anti-DNAJC19 (1:2000 dilution antibody; both from Sigma Aldrich), anti-NOB1 and anti-PES1 (1:1000 dilution; Sigma Aldrich) Tubulin and β-actin antibody (1:5000 dilution; Abcam) was used as a loading control. Horseradish-peroxidase-linked goat anti-rabbit IgG (1:5000 dilution; Santa-Cruz Biotechnology) was used as a secondary antibody. Blots were developed with enhanced chemiluminescence (ECL). Densitometry analysis was performed using BioRad ImageLab software.
Rabbit polyclonal anti oxphos
Manufactured by Abcam
Sourced in United Kingdom
The Rabbit polyclonal anti-OXPHOS is a primary antibody that recognizes the oxidative phosphorylation (OXPHOS) complexes in mammalian cells. It can be used to detect the presence and levels of these complexes in various sample types, including cell lysates and tissue extracts.
Automatically generated - may contain errors
2 protocols using rabbit polyclonal anti oxphos
1
Protein Extraction and Immunoblot Analysis
2
DIMT1 and OXPHOS Protein Quantification
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Cells were washed with phosphate-buffered saline (PBS) and homogenized in cell lysis buffer (150mM NaCl, 1% NP40, 10% DOC, 10% SDS, 50mM Tris, pH 7.4). Determination of protein concentration was performed using the BCA protein assay kit (Pierce, Rockford, IL). Samples were mixed with standard Laemmli loading buffer and 30 µg protein loaded on mini-Protean TGX precast gel (Bio-Rad, USA). After SDS-PAGE, proteins were transferred to Immunoblot PVDF membranes (BioRad), the membrane blocked with 3% BSA, and further incubated with rabbit monoclonal anti-DIMT1 antibody (1:1000 dilution) and Rabbit polyclonal anti-OXPHOS (1:250 dilution antibody; both from Abcam, Cambridge, UK). Tubulin antibody
(1:5000 dilution; Abcam, Cambridge, UK) was used as a loading control. Horseradish peroxidase-linked goat anti-rabbit IgG (1:5000 dilution; Santa-Cruz Biotechnology) was used as secondary antibody. Blots were developed with enhanced chemiluminescence (ECL).
Densitometry analysis was performed using BioRad ImageLab software.
(1:5000 dilution; Abcam, Cambridge, UK) was used as a loading control. Horseradish peroxidase-linked goat anti-rabbit IgG (1:5000 dilution; Santa-Cruz Biotechnology) was used as secondary antibody. Blots were developed with enhanced chemiluminescence (ECL).
Densitometry analysis was performed using BioRad ImageLab software.
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