Fig 1.
Specificity of novel tau antibodies to the MT-binding repeat domains of tau.
Immunoblots of recombinant glutathione-S-transferase (GST), GST/R1-3, GST/R1, GST/R3 and GST/R4 human tau probed with anti-GST antibody and the novel tau antibodies (as indicated for each blot) 81A11, 83E4, 94-3A2, 94-3A6, 94-4F1 to determine epitopes. The mobilities of molecular mass markers are shown on the left.
Fig 2.
Specificity of novel tau antibodies to recombinant 3-repeat and 4-repeat human tau proteins.
Immunoblots of recombinant K18 tau fragment (4R MTBD) and full-length human 2N3R and 2N4R tau were probed with the novel tau antibodies (as indicated for each blot) 81A11, 83E4, 94-3A2, 94-3A6, 94-4F1 to determine specificity for 3R and 4R tau. The mobilities of molecular mass markers are shown on the left.
Fig 3.
Characterization of novel tau antibodies in NTg, tau KO and PS19 tau Tg mice.
Immunoblots of total brain lysates from NTg, PS19 tau transgenic and tau KO (-/-) mice were probed with the novel tau antibodies (as indicated for each blot) 81A11, 83E4, 94-3A2, 94-3A6, 94-4F1 to determine affinity for tau in mice. An immunoblot probed with secondary antibody only highlights a non-specific band present on all immunoblots (arrow). The mobilities of molecular mass markers are shown on the left.
Fig 4.
Immunohistological identification of tau inclusions in JNPL3 tau Tg mice and PS19 tau Tg mice.
Immunohistochemistry of representative tau pathology in the spinal cord of 7.5 month old JNPL3 Tg mice and 13 month old PS19 Tg mice detected with the novel tau antibodies 81A11, 83E4, 94-3A2, 94-3A6, 94-4F1. A 7 month old NTg mouse and 6 month old tau KO (Tau-/-) mouse are also shown. Scale bar = 100 μm.
Fig 5.
Characterization of novel tau antibodies in sarkosyl-insoluble human control and AD cortex.
Immunoblots of biochemically sarkosyl-insoluble temporal cortex lysates from post-mortem control (n = 2) and AD (n = 3) cases were probed with the novel tau antibodies (as indicated for each blot) 81A11, 83E4, 94-3A2, 94-3A6, 94-4F1 to determine affinity for tau in human AD. The mobilities of molecular mass markers are shown on the left.
Fig 6.
Immunohistological reactivity of tau inclusions in human post-mortem AD cortex and hippocampus and PSP basal ganglia.
Immunohistochemistry of representative tau pathology in post-mortem AD and control temporal cortex and hippocampus and in PSP and control basal ganglia detected with the novel tau antibodies 81A11, 83E4, 94-3A2, 94-3A6, 94-4F1. Asterisks mark astroglial tau inclusions and arrows mark NFTs. Scale bar = 100 μm.
Table 1.
Novel MTBD tau antibodies generated in this study.
Summary of the new antibodies generated and characterized in this study, including the antigens used to generate each antibody, the specificity for each antibody, and their isotypes.