Figure 2.

mAb 1E4 recognizes a modified epitope on Npl3p. Duplicate immunoblots were probed with anti-Npl3p mAbs 2F1 (A) or 1E4 (B). Lanes 1–3 and 7–9 contain 0.25, 0.5, or 1.0 μl of whole-cell yeast extract (approximately 10 mg of total protein per ml), respectively. Lanes 4–6 and 7–9 contain 12.5, 25, or 50 ng of recombinant Npl3p (rNpl3p) expressed in bacteria and purified exploiting a six-histidine tag that also increases the molecular mass of the protein relative to untagged Npl3p expressed in yeast (yNpl3p).