Observed band at 18 kDa: LC3-I.
Observed band at 17 kDa: LC3-II.
Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody (ab48394)This image is courtesy of an Abreview by Armen Petrosyan.
ab48394 staining LC3B in a Rat hepatocyte by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.2% Triton X-100 in PBS and blocked with 1% Donkey serum in 0.1% PBST for 60 minutes at 21°C. Samples were incubated with primary antibody (1/50 in PBS + 1% BSA) for 3 hours at 22°C. An Alexa Fluor® 394-conjugated Donkey anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/200.
ab48394 staining LC3B (LC3-II) in HeLa cells treated with calmidazolium chloride (ab120658), by ICC/IF. Increase of LC3B (LC3-II) expression correlates with increased concentration of calmidazolium chloride, as described in literature. The cells were incubated at 37°C for 6h in media containing different concentrations of ab120658 (calmidazolium chloride) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab48394 (1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells with Dylight 488 (green).Nuclei and alpha -tubulin were counterstained with DAPI (blue) and Dylight 550 (red).
Western blot - Anti-LC3B antibody (ab48394)This image is courtesy of an anonymous Abreview
All lanes : Anti-LC3B antibody (ab48394) at 1/2000 dilution
Lane 1 : Rat whole tissue lysate - Normal liver Lane 2 : Rat whole tissue lysate - liver treated with AEE788 at 50 mg/kg 3 times a week for 1 week Lane 3 : Rat whole tissue lysate - liver treated with RAD at 2.5 mg/kg daily for 1 week
Lysates/proteins at 30 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution