The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use a concentration of 0.5 µg/ml. Predicted molecular weight: 53 kDa.
This is a component of the ubiquinol-cytochrome c reductase complex (complex III or cytochrome b-c1 complex), which is part of the mitochondrial respiratory chain. This protein may mediate formation of the complex between cytochromes c and c1.
Belongs to the peptidase M16 family. UQCRC1/QCR1 subfamily.
Cytochrome b c1 complex subunit 1, mitochondrial antibody
Cytochrome b-c1 complex subunit 1 antibody
Ubiquinol cytochrome c reductase core protein I antibody
Ubiquinol-cytochrome-c reductase complex core protein 1 antibody
Immunocytochemistry/ Immunofluorescence - Anti-Ubiquinol-Cytochrome C Reductase Core Protein I antibody [16D10AD9AH5] (ab110252)
ab110252 stained HCT116 cells. The cells were 100% methanol fixed for 5 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab110252 at 1/50 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150117 used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Western blot - Anti-Ubiquinol-Cytochrome C Reductase Core Protein I antibody [16D10AD9AH5] (ab110252)
All lanes : Anti-Ubiquinol-Cytochrome C Reductase Core Protein I antibody [16D10AD9AH5] (ab110252) at 0.5 µg/ml
Lane 1 : Human heart mitochondrial lysate Lane 2 : Cow heart mitochondrial lysate Lane 3 : Rat heart mitochondrial lysate Lane 4 : Mouse heart mitochondrial lysate Lane 5 : HepG2 mitochondrial lysate
Predicted band size : 53 kDa Extra bands in the Mouse sample (lane 4) are due to the reaction of the IgG-specific goat anti-mouse secondary antibody with residual mouse blood in the heart tissue, as it is very difficult to entirely remove the blood from these small organs.
Flow Cytometry - Anti-Ubiquinol-Cytochrome C Reductase Core Protein I antibody [16D10AD9AH5] (ab110252)
Overlay histogram showing HepG2 cells stained with ab110252 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab110252, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Kazak L et al. UCP1 deficiency causes brown fat respiratory chain depletion and sensitizes mitochondria to calcium overload-induced dysfunction. Proc Natl Acad Sci U S A114:7981-7986 (2017).
Read more (PubMed: 28630339) »
Hernansanz-Agustín P et al. Mitochondrial complex I deactivation is related to superoxide production in acute hypoxia. Redox Biol12:1040-1051 (2017).
Read more (PubMed: 28511347) »