Mouse anti human phf tau at8

One hundred pHuC-h4R-tau-GFP-injected embryos or wild-type embryos were collected at 24 hpf for independent experiments. The sample preparation was based on the protocols described in the Zebrafish Book [34 ]. Clarified lysates containing equivalent amounts of proteins derived from zebrafish embryos were analyzed by Western blotting with the following antibodies: mouse anti-human PHF-Tau (AT8) (1:3000) (Thermo Fisher Scientific), rabbit anti-GFP (1:5000) (Abcam), mouse anti-GAPDH (1D4) (1:5000) (Novus Biologicals, Inc., Littleton, CO,USA), HRP-conjugated AffiniPure goat anti-mouse IgG (1:20000) (Jackson ImmunoResearch Laboratories, Inc., West Grove, PA).

Whole-mount immunostaining was performed following standard protocols as previously described [41 (link)] with some modifications. The antibodies used were as follows: mouse anti-GFP (1E4) (1:300) (MBL, Nagoya, Japan), mouse anti-HA (F-7) (1:100) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), mouse anti-human PHF-Tau (AT8) (1:100) (Thermo Fisher Scientific, West Palm Beach, FL), rabbit anti- Caspase-9 (Novus Biologicals, Inc., Littleton, CO,USA), rabbit anti-Tau [pT212] (Thermo Fisher Scientific), rabbit anti-GFP (1:300) (Abcam, Cambridge, UK), Cy3-conjugated anti-mouse IgG (1:100), Cy2-conjugated anti-mouse IgG (1:100), Cy2-conjugated anti-rabbit IgG (1:100) and Cy3-conjugated anti-rabbit IgG (1:100) (Jackson ImmunoResearch Laboratories, Inc., West Grove, PA). High resolution images of the samples were captured using a Leica SP5 X Inverted Confocal Microscope.