Tau is a microtubule-associated phosphoprotein (MAP) localized in neuronal axons. It promotes tubulin polymerization and stabilizes microtubules. Tau proteins constitute a family of six isoforms ranging in size from 352 to 441 amino acids. The tau variants differ from each other in the presence or absence of either three or four repeat-regions near the carboxy-terminal and the presence or absence of one or two inserts near the amino-terminal. Tau is hyperphosphorylated by ERK, GSK-3, TPKII, and CDK5. Over thirty phosphorylation sites have been described, including Thr39, Ser46, Thr50, Thr69, Thr153, Thr175, Thr181, Ser198, Ser199, Ser202, Thr205, Ser208, Ser210, Thr212, Ser214, Thr217, Thr231, Ser235, Ser237, Ser241, Ser262, Ser285, Ser305, Ser324, Ser352, Ser356, Ser396, Ser400, Thr403, Ser404, Ser409, Ser412, Ser413, Ser416, and Ser422. Specifically, TPKII phosphorylates serines 202 and 404. GSK-3β transfection phosphorylates serines 199, 202, 235, 396, 404, and 413 and threonines 205 and 231. These sites are among the major abnormal phosphorylation sites of Tau. Phosphorylation on these sites reduces the ability of a given tau species to promote microtubule self-assembly. Hyperphosphorylated tau is the major protein found in the paired helical filaments (PHFs), which make up the pathological neurofibrillary tangles of Alzheimer’s disease (AD). These PHFs are also found in the lesions of other central nervous system disorders. GenScript Rabbit Anti-Tau (Phospho-Ser235) Polyclonal Antibody is developed in rabbit using a KLH-coupled synthetic phosphopeptide corresponding to the residues surrounding serine 235 of human tau.
Affinity chromatography using phospho-peptide, and cross-adsorbed with the corresponding non-phosphopeptide
Reconstitute the lyophilized powder with deionized water (or equivalent) to an antibody concentration of 0.5 mg/ml.
GenScript Rabbit Anti-Tau (Phospho-Ser235) Polyclonal Antibody detects endogenous levels of tau protein only when phosphorylated at serine 235.
The antibody is stable in lyophilized form if stored at -20°C or below. The reconstituted antibody can be stored for 2-3 weeks at 2-8°C. For long term storage, aliquot and store at -20°C or below. Avoid repeated freezing and thawing cycles.
Small volumes of antibody will occasionally become entrapped in the seal of the product vial during shipment and storage. Before use, briefly centrifuge the vial to dislodge any liquid in the container's cap.
Working concentrations for specific applications should be determined by the investigator. The appropriate concentrations may be affected by secondary antibody affinity, antigen concentration, the sensitivity of the method of detection, temperature, the length of the incubations, and other factors. The suitability of this antibody for applications other than those listed below has not been determined. The following concentration ranges are recommended starting points for this product. ELISA: 0.05-0.2 µg/ml Western blot: 0.5-2 µg/ml Other applications: user-optimized
Western blot analysis of mouse brain tissue lysate using 2 µg/ml Rabbit Anti-Tau (Phospho-Ser235) Polyclonal Antibody (GenScript, A01096) Lane 1: Rabbit Anti-Tau (Phospho-Ser235) Polyclonal Antibody Lane 2: Rabbit Anti-Tau (Phospho-Ser235) Polyclonal Antibody pre-incubated with non-phospho-peptide Lane 3: Rabbit Anti-Tau (Phospho-Ser235) Polyclonal Antibody pre-incubated with phospho-peptide Lane 4: Rabbit Anti-Tau (Phospho-Ser235) Polyclonal Antibody pre-incubated with generic phospho-serine containing peptide The signal was developed with IRDyeTM 800 Conjugated Goat Anti-Rabbit IgG.