Figure 2

Western Blot analysis of RAb^DMvIII antibody. A) 30 ug of U87vIII and U87 wild type cell lysate loaded under reducing conditions on 4-20% SDS-PAGE. A) RAb^vIII recognizes 145 kDa band which corresponds to the EGFRvIII and 45 kDa unspecific protein band. A) RAb^DMvIII recognizes only 145 kDa protein band. B) RAb^DMvIII recognizes 170 KDa protein band in 30 ug of A431 cell lysate (cells overexpressing EGFR) and 145 kDa band in 30 ug of HC2 cell lysate under reducing conditions. B) EGFR antibody recognizes equal amount of protein in both HC2 and A431 cell lysate under reducing conditions. C) RAb^DMvIII under non reducing conditions detect only 145 kDa protein band whereas RAb^vIII still recognizes 145 kDa and 170 kDa protein bands in non reducing conditions. C) Actin antibody under reducing condition shows equal amount of protein loaded in the wells.