Western blot analysis of extracts from various tissues, untreated (-) or λ phosphatase-treated (+), using Phospho-Tau (Ser46) Rabbit Antibody (upper), Tau (D1M9X) XP® Rabbit mAb #46687 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower). Tau-KO mouse brains were kindly provided by Dr. Dominic Walsh at Brigham and Women's Hospital and Harvard Medical School.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Phospho-Tau (Ser46) Rabbit Antibody recognizes endogenous levels of Tau protein only when phosphorylated at Ser46.
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser46 of human Tau protein. Antibodies are purified by peptide affinity chromatography.
Tau is a heterogeneous microtubule-associated protein that promotes and stabilizes microtubule assembly, especially in axons. Six isoforms with different amino-terminal inserts and different numbers of tandem repeats near the carboxy terminus have been identified, and tau is hyperphosphorylated at approximately 25 sites by Erk, GSK-3, and CDK5 (1,2). Phosphorylation decreases the ability of tau to bind to microtubules. Neurofibrillary tangles are a major hallmark of Alzheimer's disease; these tangles are bundles of paired helical filaments composed of hyperphosphorylated tau. In particular, phosphorylation at Ser396 by GSK-3 or CDK5 destabilizes microtubules. Furthermore, research studies have shown that inclusions of tau are found in a number of other neurodegenerative diseases, collectively known as tauopathies (1,3).
Phosphorylation of Tau at Ser46 is induced by different kinases, including GSK-3β (4), casein kinase-1 (5), and p38MAPK (6). Tau phosphorylation at Ser46 has been linked to different neurodegenerative diseases, including in PHF-Tau from Alzheimer's disease (4,6), Progressive Supranuclear Palsy (7), and synucleinopathies (6).
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