NeuroMab™ Anti-pTau BBB Shuttle Antibody(NRZP-1022-ZP3068)

Figure 1: Accumulation of oligomeric tau depends on yeast growth characteristics.

A pho85Δ yeast strain transformed with a HIS6-PG-TEV-hTau (2N/4R) plasmid was inoculated in SD-URA medium. Subsequent sampling at different OD's was followed by protein extraction and non-reducing SDS-PAGE. Tau was detected with the ADx215 monoclonal antibody. Besides monomeric tau (∼75 kDa), higher weight dimeric and higher oligomeric species of tau can be detected.

Figure 2 Selectivity of tau aggregates and phosphorylation dependence of anti-tau monoclonal antibodies.

A) Western blot analysis of recombinant HIS-tagged tau obtained from E. coli and protein extracts obtained from control and humanized yeast strains using the commercial pan-tau antibody Tau-5 and ADx210 mAb as indicated. The number between brackets refers to the exposure time. The solid bar refers to high molecular weight and oligomeric tau and the small arrow in the picture of ADx210 to a presumable tau dimer. B) Western blot analysis of purified tau obtained from the pho85Δ strain before and after treatment with shrimp alkaline phosphatase (SAP) following different time intervals as indicated. Recombinant HIS-tagged tau obtained from E. coli served as control. The pan-tau antibody Tau-5, the phosphorylation-specific antibody AT270, and the antibody BT2 recognizing non-phosphorylated tau, were used in addition to the novel mAbs ADx215 and ADx210.

Figure 3: Analysis of novel mAbs on transgenic mouse and human brain samples.

Western blot analysis of brain extracts from control mice (FVB), knock-out mice and transgenic (Tg) mice expressing a clinical mutant of human tau (Tau-P301L). The age of the mice is indicated between brackets. The arrow on the second blot indicates high molecular weight tau complexes. The antibodies used are ADx215 (YT1.15), ADx210 (YT1.10) and the commercial phosphorylation-specific antibody AT100 (exposure time of 30 min for ADx210 (YT1.10) and AT100, and 30 sec for ADx215 (YT1.15)).

Figure 4: Analysis of novel mAbs on transgenic mouse and human brain samples.

Immunohistochemical analysis of different regions of the Tau-P301L Tg mouse using ADx215 (YT1.15) and ADX210 (YT1.10) for immunodetection.

Figure 5: Immunodetection of aggregated tau from pho85d yeast in ELISA using pan-tau antibody.