The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 48 kDa (predicted molecular weight: 44 kDa).
FunctionCysteine protease required for autophagy, which cleaves the C-terminal part of either MAP1LC3, GABARAPL2 or GABARAP, allowing the liberation of form I. A subpopulation of form I is subsequently converted to a smaller form (form II). Form II, with a revealed C-terminal glycine, is considered to be the phosphatidylethanolamine (PE)-conjugated form, and has the capacity for the binding to autophagosomes.
Tissue specificityMainly expressed in the skeletal muscle, followed by brain, heart, liver and pancreas.
Sequence similaritiesBelongs to the peptidase C54 family.
Lane 1: Wild type HAP1 whole cell lysate (20 µg) Lane 2: ATG4B knockout HAP1 whole cell lysate (20 µg) Lane 3: HeLa whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab72878 observed at 68 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab72878 was shown to recognize ATG4B when ATG4B knockout samples were used, along with additional cross-reactive bands. Wild-type and ATG4B knockout samples were subjected to SDS-PAGE. ab72878 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - ATG4B antibody (ab72878)
All lanes : Anti-ATG4B antibody (ab72878) at 1 µg/ml
Lane 1 : Human heart tissue lysate - total protein (ab29431) Lane 2 : Human liver tissue lysate - total protein (ab29889)
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution Developed using the ECL technique