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Up-regulation and Antibody-Peptide Competition. Human recombinant Tau stimulated with GSK-3 beta; (1 µg per µg Tau) for 45 minutes was added to background extracts, resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature and either left untreated (1-4) or treated with lambda phosphatase (5), then incubated with phospho-Tau [pS422] polyclonal antibody (Product # OPA1-03151) in a 3% BSA-TBST buffer for two hours at room temperature, following prior incubation with: no peptide (1,5), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphoserine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F(ab')2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ method.The data show that only the phosphopeptide corresponding to Tau [pS422] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phosphopeptide derived from a region of human Tau that contains serine 422.|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||Assay-Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
OPA1-03151 detects human p-Tau (pSer422) from human samples. Mouse and rat share 100% homology with the immunizing peptide but have not yet been tested.
OPA1-03151 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~63 kDa protein representing p-Tau (pSer422).
OPA1-03151 immunizing phosphopeptide was derived from a region of human tau that contains serine 422.
For up to 1 week, store at 4°C. For long-term storage, store -20°C.
Tau is a neuronal microtubule-associated protein found predominantly on axons and functions to promote tubulin polymerization and stabilize microtubules. Tau, in its hyperphosphorylated form, is the major component of paired helical filaments (PHF), the building block of neurofibrillary lesions in Alzheimer's disease (AD) brain. Hyperphosphorylated Tau is also found in neurofibrillary lesions in a range of other central nervous system disorders. Hyperphosphorylation impairs the microtubule binding function of Tau, resulting in the destabilization of microtubules in AD brains, ultimately leading to the degeneration of the affected neurons. Numerous serine/threonine kinases, includingGSK-3beta, protein kinase A (PKA), cyclin-dependent kinase 5 (cdk5) and casein kinase II (CK2), phosphorylate Tau. Serine 422 can be phosphorylated by members of the MAP kinase (MAPK) family.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
DDPAC; Disinhibition Dementia Parkinsonism Amyotrophy Complex; FLJ31424; FTDP17; G protein beta1/gamma2 subunit-interacting factor 1; MAPTL; Microtubule Associated Protein Tau; microtubule-associated protein tau; microtubule-associated protein tau, isoform 4; MSTD; Neurofibrillary Tang; neurofibrillary tangle protein; paired helical filament-tau; PHF-tau; protein phosphatase 1, regulatory subunit 103; Tau microtubule-associated protein
AI413597; AW045860; DDPAC; FTDP-17; MAPT; MAPTL; MSTD; Mtapt; MTBT1; MTBT2; PPND; PPP1R103; pTau; RNPTAU; TAU