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 70 kDa (predicted molecular weight: 60 kDa).
Thrombomodulin is a specific endothelial cell receptor that forms a 1:1 stoichiometric complex with thrombin. This complex is responsible for the conversion of protein C to the activated protein C (protein Ca). Once evolved, protein Ca scissions the activated cofactors of the coagulation mechanism, factor Va and factor VIIIa, and thereby reduces the amount of thrombin generated.
Endothelial cells are unique in synthesizing thrombomodulin.
Involvement in disease
Defects in THBD are the cause of thrombophilia due to thrombomodulin defect (THR-THBD) [MIM:188040]. A hemostatic disorder characterized by a tendency to thrombosis. Defects in THBD are a cause of susceptibility to hemolytic uremic syndrome atypical type 6 (AHUS6) [MIM:612926]. An atypical form of hemolytic uremic syndrome. It is a complex genetic disease characterized by microangiopathic hemolytic anemia, thrombocytopenia, renal failure and absence of episodes of enterocolitis and diarrhea. In contrast to typical hemolytic uremic syndrome, atypical forms have a poorer prognosis, with higher death rates and frequent progression to end-stage renal disease. Note=Susceptibility to the development of atypical hemolytic uremic syndrome can be conferred by mutations in various components of or regulatory factors in the complement cascade system. Other genes may play a role in modifying the phenotype.
Western blot - Anti-Thrombomodulin antibody (ab94373)
All lanes : Anti-Thrombomodulin antibody (ab94373) at 1 µg/ml
Lane 1 : Human heart tissue lysate - total protein (ab29431) Lane 2 : Lung (Human) Tissue Lysate Lane 3 : Human kidney tissue lysate - total protein (ab30203) Lane 4 : Human placenta tissue lysate - total protein (ab29745)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Thrombomodulin contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.