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: PARP1 knockout HAP1 whole cell lysate (20 µg) Lane 3: HeLa whole cell lysate (20 µg) Lane 4: MCF7 whole cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab72878 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab72878 was shown to recognize PARP1 when PARP1 knockout samples were used, along with additional cross-reactive bands. Wild-type and PARP1 knockout samples were subjected to SDS-PAGE. Ab72878 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution 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