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. 2000 Feb;11(2):703–719. doi: 10.1091/mbc.11.2.703

Figure 1.

Figure 1

Characterization of anti-NTF2 mAbs. (A) Anti-NTF2 mAbs are monospecific for NTF2. Recombinant human untagged (15 ng) or myc-tagged (85 ng) NTF2 protein and HeLa cytosol were separated on 15% SDS-PAGE gels, transferred to nitrocellulose, and probed with anti-NTF2 mAbs. 5A3, 7E5, 2C1, and 4F5 were used at 5 μg/ml purified antibody. 5E8 was used at 1:5 dilution of tissue culture supernatant. (B) Epitope mapping of anti-NTF2 mAbs. ELISAs were performed using recombinant wild-type human NTF2, recombinant mutant human NTF2 (D117N, H124Δ, and F126Δ), and recombinant wild-type yeast NTF2. (C) Assignment of the epitope of anti-NTF2 5E8. Western blot of recombinant untagged human NTF2 (15 ng), HeLa cytosol, and Xenopus lysate probed with 5E8 (1:1000 of ascites fluid). The synthetic N-terminal peptide of human NTF2 used to generate 5E8 is shown above the corresponding N-terminal 14 amino acids from Xenopus NTF2. Residues conserved between human and Xenopus NTF2 are in bold. The bracket indicates residues that likely encompass the 5E8 epitope.