The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/10000 - 1/50000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
May be involved in macrophage-mediated cellular proliferation. It is mitogenic for fibroblasts and smooth muscle but not endothelial cells. It is able to bind EGF receptors with higher affinity than EGF itself and is a far more potent mitogen for smooth muscle cells than EGF. Also acts as a diphtheria toxin receptor.
Contains 1 EGF-like domain.
Several N-termini have been identified by direct sequencing. The forms with N-termini 63, 73 and 74 have been tested and found to be biologically active. O-linked glycan attachment sites were determined by Edman degradation, O-glycanase digest suggests mucin-type glycosylation (done in HB-EGF purified from histiocytic lymphoma cell line U-937).
Cell membrane and Secreted > extracellular space. Mature HB-EGF is released into the extracellular space and probably binds to a receptor.
Western blot analysis on immunoprecipitation from 1) K562 cell lysate and 2) PBS, labeling HB EGF using ab185555 at 1/50 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG at a 1/1500 dilution.
Western blot - Anti-HB EGF antibody [EPR13276(2)] (ab185555)
All lanes : Anti-HB EGF antibody [EPR13276(2)] (ab185555) at 1/20000 dilution
Lane 1 : HeLa cell lysate Lane 2 : K562 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 23 kDa Observed band size: 23 kDa