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KO validation (Figure 1 and 2): Anti-ADAM10 antibody (2051) specificity was further verified by ADAM10 specific knockout. ADAM10 signal in was detected by 2051 in wild type of MEF cells and 293 cells, but not in ADAM10 knockout cells.
Independent Antibody Validation in Cell lines (Figure 3) shows similar ADAM10 expression profile in human and mouse cell lines detected by two independent anti-ADAM10 antibodies that recognize different epitopes, 2051 against C-terminus domain and 24-024 against human recombinant fusion protein. ADAM10 proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies.
Regulated expression validation (Figure 12 and 14): ADAM10 expression detected by anit-ADAM10 antibodies (2051) was down-regulated when exposed to oxygen-glucose deprivation (OGD) or glutamate (Fig 8), and up-regulated in membrane compartment whereas down-regulated in intracellular fractions when treated with donepezil (Fig 14).
KD validation (Figure 15): Anti-ADAM10 antibody (2051) specificity was further verified by ADAM10 specific knockdown. ADAM10 signal in HEK/APP cells transfected with ADAM10 siRNA or Ferrochelatase siRNA (F) or N-methylprotoporphyrin IX siRNA (N) was disrupted in comparison with that in mock-transfected cells (M) or cells transfected with TACE siRNA (T).
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Daudi Lysate
ADAM10 Peptide
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