The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/300. PubMed: 17021035
1/5000. Predicted molecular weight: 50-55 kDa. This dilution was sufficient for detection of Cytochrome P450 in 20ug of human liver microsomes by colorimetric immunoblot analysis using Goat anti-Rabbit IgG:AP as the secondary antibody.
Use at an assay dependent concentration. PubMed: 24155903
Metabolizes several precarcinogens, drugs, and solvents to reactive metabolites. Inactivates a number of drugs and xenobiotics and also bioactivates many xenobiotic substrates to their hepatotoxic or carcinogenic forms.
Western blot - Anti-Cytochrome P450 2E1 antibody (ab28146)
All lanes : Anti-Cytochrome P450 2E1 antibody (ab28146) at 1/1000 dilution
Lane 1 : Molecular weight marker Lane 2 : Cell lysates prepared from human liver microsomes Lane 3 : Cell lysates prepared from rat liver microsomes Lane 4 : Cell lysates prepared from mouse liver microsomes Lane 5 : Cell lysates prepared from rabbit liver microsomes
Predicted band size: 50-55 kDa
Immunohistochemistry (Frozen sections) - Anti-Cytochrome P450 2E1 antibody (ab28146)This image is courtesy of an anonymous Abreview
ab28146 staining cells of rat liver sections by IHC-Fr. Cells were fixed in acetone and blocked with 1% BSA for 5 minutes at 25°C prior to incubating with ab28146 diluted 1/300 for 30 minutes at 25°C. An Alexa Fluor® 594 conjugated goat anti-rabbit antibody was used as the secondary.
Double IF labeling with CYP2E1 (Red) and dipeptidyl peptidase IV (Green)
ICC/IF image of ab28146 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 (ab28146, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.
Western blot - Anti-Cytochrome P450 2E1 antibody (ab28146)Image courtesy of an anonymous Abreview.
All lanes : Anti-Cytochrome P450 2E1 antibody (ab28146) at 1/2500 dilution
All lanes : Tissue lysate prepared from normal murine liver
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat anti-rabbit IgG-HRP at 1/5000 dilution
Hao L et al. Mitochondria-targeted ubiquinone (MitoQ) enhances acetaldehyde clearance by reversing alcohol-induced posttranslational modification of aldehyde dehydrogenase 2: A molecular mechanism of protection against alcoholic liver disease. Redox Biol14:626-636 (2018).
Read more (PubMed: 29156373) »
Donepudi AC et al. Deficiency of cholesterol 7a-hydroxylase in bile acid synthesis exacerbates alcohol-induced liver injury in mice. Hepatol Commun2:99-112 (2018).
Read more (PubMed: 29404516) »