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. Author manuscript; available in PMC: 2012 Apr 6.
Published in final edited form as: J Alzheimers Dis. 2012;28(2):433–441. doi: 10.3233/JAD-2011-111307

Figure 2.

Figure 2

a) NEP2 detected in blood cells by endpoint PCR. NEP2 detected in human peripheral blood mononuclear cells (PBMC) of three NI individuals using qRT-PCR NEP2-specific primers. NEP2-expressing 293T (293) cell lysate was used as a positive control and mock reverse transcription (−) was used as a negative control. b) Immunoprecipitation/Western blot hybridization (IP-Western) of NEP2-β and NEP2-γ. Western blot hybridization of NEP2-β, NEP2-γ, and GFP transfected 293T cell lysates (rows 1–3) and NEP2-β, NEP2-γ, and GFP transfected 293T cell lysates (rows 4–6) immunoprecipitated with NEP2-specific antibody (AF2340). NEP2-β and NEP2-γ were both detected in the cell lysate (rows 1–2) and were both present at higher levels following immunoprecipitation (rows 4–5).

c) Recombinant human NEP2 (rhNEP2) immunoprecipitation activity assay. rhNEP2 shows no remaining activity following AF2340 immunoprecipitation validating the specificity of the AF2340 antibody to NEP2. Samples were run in triplicate (n=3). **p<0.01. d) Recombinant human NEP (rhNEP) immunoprecipitation activity assay. rhNEP remains fully active following AF2340 and SN5c immunoprecipitation indicating that there is no cross reactivity of rhNEP to NEP2-specific and GFP-specific antibodies. Samples were run in triplicate (n=3).