Anti-Cytochrome P450 2E1 antibody (ab28146)

Immunohistochemistry (Frozen sections) - Cytochrome P450 2E1 antibody (ab28146)

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)

This image is courtesy of an anonymous Abreview

Immunocytochemistry/ Immunofluorescence - Cytochrome P450 2E1 antibody (ab28146)

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 +4ºC. 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 - 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

Western blot - Cytochrome P450 2E1 antibody (ab28146)

All lanes : Anti-Cytochrome P450 2E1 antibody (ab28146) at 1/2500 dilution

Lane 1 : Tissue lysate prepared from normal murine liver
Lane 2 : Tissue lysate prepared from normal murine liver
Lane 3 : Tissue lysate prepared from normal murine liver
Lane 4 : Tissue lysate prepared from normal murine liver

Lysates/proteins at 10 µg per lane.

Secondary
Goat anti-rabbit IgG-HRP at 1/5000 dilution
developed using the ECL technique

Predicted band size : 50-55 kDa
Observed band size : 55 kDa (why is the actual band size different from the predicted?)


Exposure time : 3 seconds

Image courtesy of an anonymous Abreview.