Detection of the 60-kDa band is insensitive to blocking of the antisera with specific inhibitor-TP.

<p>(A) Schematic representation of immunogen-α and the corresponding inhibitor-TPα. Pink shading outlines the epitope of interest. (B) Lysates of differentiated N2a cells were treated with 100 µM GSNO and then subjected to the biotin switch protocol. The resulting biotinylated lysates were fractionated by non-reducing SDS-PAGE and transferred to a nitrocellulose membrane. Following blocking the membrane was cut into stripes and each stripe was then probed with the indicated antiserum (<i>AS</i>) diluted at 1∶500 pre-incubated in the presence (<i>+ inh-TP</i>) or absence of inhibitor-TP. Successful application of the biotin switch protocol was confirmed with anti-biotin antibodies (<i>anti-biotin</i>) or streptavidin-HRP (<i>streptavidin</i>) and the position of tubulin bands was located with anti-β-tubulin antibodies (<i>β-tubulin</i>) as indicated.