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| | Tau Protein Assay |
| Description: | The Tau content of fractions was determined by dot blot analysis using nitrocellulose membranes (0.2 μm porosity) as described previously [Cisek et al., Biophys. Chem., 170:25-33]. The membranes were blocked in 5% nonfat dry milk dissolved in blocking buffer (100 mM Tris-HCl, pH 7.4, and 150 mM NaCl) for 1 h and then incubated with primary antibody Tau5 at 1:5000 dilution for 2 h. Membranes were then washed three times with blocking buffer and incubated with horseradish peroxidase-linked secondary antibody for 2 h. After washing another three times with blocking buffer, membranes were imaged using the Enhanced Chemiluminescence Western blotting Analysis System (GE Healthcare, Buckinghamshire, UK) recorded on an Omega 12iC Molecular Imaging System and quantified using Ultra-Quant software (UltraLum, Claremont, CA, USA). |
| Affinity data for this assay | |
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