Figure 2

Peptide antibodies specifically recognize the MDGA2 protein. (A) Structural representation of the MDGA2 protein, with the different domains and amino acids chosen for immunization indicated. Immunoglobulin (Ig) domains are represented as open circles, the fibronectin type III repeat (FN3) is depicted as a box and the MAM domain is shown as a hexagonal structure. Two of the peptides used for immunization are located within Ig domains, the third peptide was generated against an amino acid sequence deriving from the MAM domain. (B) Demonstration of MDGA2 antibody specificity. Recombinant flag-tagged MDGA2, MDGA1, NrCAM and axonin-1 were separated by SDS-PAGE and the different proteins were detected by western blotting using either a monoclonal antibody against the flag tag or the newly generated MDGA2 peptide antibodies. While the flag antibody clearly recognizes all recombinant IgCAMs, the pooled MDGA2 peptide antibodies specifically recognize the MDGA2 protein. Note that even when loading higher protein concentrations or using much longer exposure times, no cross-reactivity of the MDGA2 peptide antibodies with other IgCAMs could be detected.