The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 10 µg/ml.
Use a concentration of 1 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 31 kDa (predicted molecular weight: 31 kDa).
Ferric-chelate reductase that reduces Fe(3+) to Fe(2+). Present at the brush border of duodenal enterocytes where it probably reduces dietary Fe(3+) thereby facilitating its transport into the mucosal cells. Uses ascorbate as electron donor. May be involved in extracellular ascorbate recycling in erythrocyte membranes. May also act as a ferrireductase in airway epithelial cells.
Present in erythrocyte membranes (at protein level). Also expressed in respiratory epithelium.
Western blot - Anti-Cytochrome b reductase 1 antibody (ab66048)
Anti-Cytochrome b reductase 1 antibody (ab66048) at 1 µg/ml + A20 whole cell lysate (ab7180) at 10 µg
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 31 kDa Observed band size: 31 kDa
Exposure time: 10 minutes
Immunohistochemistry (Frozen sections) - Anti-Cytochrome b reductase 1 antibody (ab66048)This image is courtesy of an abreview submitted by Ruma Raha-Chowdhury, University Of Cambridge, United Kingdom
IHC-Fr image of human duodenum section stained with ab66048. The sections were incubated in 10% normal donkey serum in 0.1% PBS- and 0.3x triton100 for 1h to permeabilise the cells and to block non-specific protein-protein interactions. The sections were then incubated with the antibody ab66048, 1µg/ml) and Anti-Actin overnight at +4°C. The secondary antibody was Alexa Alexa Fluor® 488 goat anti-rabbit IgG (H+L) and Fluor®568 donkey anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. dCytb stained apical site of duodenum and actin stained muscles and blood vessels.
Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome b reductase 1 antibody (ab66048)
ICC/IF image of ab66048 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66048, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.