Western blot analysis of extracts from COS cells, untransfected (lane 1), or MAP2C transfected (lanes 2,3), using Phospho-MAP2 (Thr1620/1623) Antibody (upper) or MAP2 Antibody #4542 (lower). In lane 3, the lysate was treated with lamda phosphatase to demonstrate phospho-specificity of the antibody. (Human MAP2C construct is a generous gift from Dr. B. Shafit-Zagardo, Dept. of Pathology, Albert Einstein College of Medicine, Bronx, NY.)
|MW (kDa)||82, 280|
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-MAP2 (Thr1620/1623) Antibody detects transfected MAP2 (at a dilution of up to 1:20,000) only when phosphorylated at threonines 1620/1623. This antibody does not cross-react with phosphorylated tau.
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr1620/1623 of human MAP2. Antibodies are purified by protein A and peptide affinity chromatography.
Microtubule-associated protein 2 (MAP2) is a neuronal phosphoprotein that regulates the structure and stability of microtubules, neuronal morphogenesis, cytoskeleton dynamics, and organelle trafficking in axons and dendrites (1). Multiple MAP2 isoforms are expressed in neurons, including high molecular weight MAP2A and MAP2B (280 and 270 kDa), and low molecular weight MAP2C and MAP2D (70 and 75 kDa). Phosphorylation of MAP2 modulates its association with the cytoskeleton and is developmentally regulated. GSK-3 and p44/42 MAP kinase phosphorylate MAP2 at Ser136, Thr1620, and Thr1623 (2,3). Phosphorylation at Thr1620/1623 by GSK-3 inhibits MAP2 association with microtubules and microtubule stability (3).
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