The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/250. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa).
FunctionCalcium/phospholipid-binding protein which promotes membrane fusion and is involved in exocytosis. This protein regulates phospholipase A2 activity. It seems to bind from two to four calcium ions with high affinity.
Sequence similaritiesBelongs to the annexin family. Contains 4 annexin repeats.
DomainA pair of annexin repeats may form one binding site for calcium and phospholipid.
Post-translational modificationsPhosphorylated by protein kinase C, epidermal growth factor receptor/kinase and TRPM7. Phosphorylation results in loss of the inhibitory activity.
Cellular localizationNucleus. Cytoplasm. Cell projection > cilium. Basolateral cell membrane. Found in the cilium, nucleus and basolateral cell membrane of ciliated cells in the tracheal endothelium (By similarity). Found in the cytoplasm of type II pneumocytes and alveolar macrophages.
Lane 1: Wild type HAP1 whole cell lysate (20 µg) Lane 2: Annexin A1 knockout HAP1 whole cell lysate (20 µg) Lane 3: HeLa whole cell lysate (20 µg) Lane 4: HepG2 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab46686 observed at 38 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab46686 was shown to specifically react with Annexin A1 when Annexin A1 knockout samples were used. Wild-type and Annexin A1 knockout samples were subjected to SDS-PAGE. ab46686 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/250 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 - Annexin A1 antibody (ab46686)
Anti-Annexin A1 antibody (ab46686) at 1/250 dilution + THP1 whole cell lysate (ab7913) at 10 µg