Invitrogen™ Phospho-Tau (Thr205) Polyclonal Antibody

Catalog No.	Quantity
44-738-G	100 μL

Catalog No. 44-738-G Supplier Invitrogen™ Supplier No. 44738G

Description

Rabbit Polyclonal Antibody

Purified from rabbit serum by sequential epitope-specific chromatography, this product contains enough material for 10 mini-blots. This antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Tau. The final product is generated by affinity chromatography using a Tau-derived peptide that is phosphorylated at threonine 205. The antibody has been used in western blotting. Previous lots of this antibody have been used in immunohistochemistry. In western analysis the positive control used was recombinant human Tau treated with GSK3beta.

Tau is a neuronal microtubule-associated protein found predominantly on axons. The function of Tau is to promote tubulin polymerization and stabilize microtubules. The C-terminus binds axonal microtubules while the N- terminus binds neural plasma membrane components, suggesting that tau functions as a linker protein between both. Axonal polarity is predetermined by TAU/MAPT localization (in the neuronal cell) in the domain of the cell body defined by the centrosome. The short isoforms allow plasticity of the cytoskeleton while the longer isoforms may preferentially play a role in its stabilization. In its hyper-phosphorylated form, Tau is the major component of paired helical filaments (PHF), the building block of neurofibrillary lesions in Alzheimer's diseases (AD) brain. Hyper-phosphorylation 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 phosphorylate Tau, including GSK-3beta, protein kinase A (PKA), cyclin-dependent kinase 5 (cdk5) and casein kinase II. Hyper-phosphorylated Tau is found in neurofibrillary lesions in a range and other central nervous system disorders such as Pick's disease, frontotemporal dementia, cortico-basal degeneration and progressive supranuclear palsy.

Specifications

• Antigen: Phospho-Tau (Thr205)
• Applications: Immunohistochemistry, Western Blot, Immunocytochemistry
• Classification: Polyclonal
• Conjugate: Unconjugated
• Formulation: Dulbecco's PBS with 1mg/mL BSA and 0.05% sodium azide; pH 7.3
• Gene: MAPT
• Gene Accession No.: P10636, P10637, P19332
• Gene Alias: AI413597; AW045860; DDPAC; FLJ31424; FTDP17; FTDP-17; G protein beta1/gamma2 subunit-interacting factor 1; map tau; Mapt; MAPTL; MGC138549; microtubule associated protein tau; microtubule-associated protein tau; microtubule-associated protein tau, isoform 4; microtubules; MSTD; Mtapt; MTBT1; MTBT2; Neurofibrillary tangle protein; neurofibrillary tangles; Neuronal Marker; paired helical filament-tau; PHFtau; PHF-tau; PPND; PPP1R103; protein phosphatase 1, regulatory subunit 103; pTau; RNPTAU; Tau; Tau microtubule-associated protein; tau protein; Tau-4; Tau5; Unknown (protein for MGC:134287)
• Gene Symbols: MAPT
• Host Species: Rabbit
• Immunogen: The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human Tau that contains threonine 205. The sequence is conserved in mouse and rat.
• Purification Method: Antigen affinity chromatography
• Quantity: 100 μL
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 17762, 29477, 4137
• Target Species: Human, Mouse, Rat
• Content And Storage: -20°C
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG

Frequently Asked Questions (FAQs)

I know that there's no single procedure that applies to all antigens, but what are some commonly used procedures for performing immunohistochemical staining of brain tissue?

Here is a procedure for the immunohistochemical staining of Beta-amyloid with paraffin embedded sections of transgenic mouse brain. The following protocol was developed for use with paraffin embedded sections stained with a variety of our antibodies against Beta-amyloid. Applicable rabbit and mouse anti-Beta-amyloid antibodies are: 51-2700, 13-0200, 71-5800, 13-0100Z, 37-4200, 43-7900, 44-136, 700254, 36-6900, and AHB0121.

Beta-amyloid Staining:
- Transgenic mice (expressing the transgenes PS1-A246E + APP swe, APP swe alone, or PS1-A246E alone) were perfused with 1 x Dulbecco's phosphate buffered saline (D-PBS) followed by 4% paraformldehyde buffered with D-PBS. Brain tissues were then embedded in paraffin prior to microtome sectioning.
- After mounting on slides, the paraffin-embedded tissue sections were then deparaffinized with heat. The sections were then incubated for 3 minutes in 70% formic acid. Next, they were deparaffinized further with xylene followed with 100% ethanol. The sections were then re-hydrated in a graded ethanol series (100% ethanol, 95% ethanol, 70% ethanol, and then water).
- Endogenous peroxidase activity was quenched with 3% hydrogen peroxide in methanol. The sections were then heated in the microwave for 5-7 minutes in water, cooled at room temperature for 5 minutes and then rinsed in water. The sections were then washed in TBS (0.05 M Tris-HCl, pH 7.6, with 0.25 M NaCl) prior to blocking. Non-specific binding was blocked with 3% normal goat serum and 0.1% Triton X-100 in TBS for 1 hour at room temperature.
- The sections were then stained with anti-Beta-amyloid antibodies. The staining solution typically consisted of the antibody at a concentration of 5 µg/mL in TBS containing 2% normal goat serum. After incubation at room temperature for at least 1 hr., the sections were washed in TBS 3 times for 5 minutes each. An anti-mouse or anti-rabbit secondary antibody labeled with HRP was then used with DAB or AEC as substrates to stain the Beta-amyloid.
[Adapted from Borchelt, D.R., et al. (1997) Accelerated amyloid deposition in the brains of transgenic mice coexpressing mutant presenilin 1 and amyloid precursor proteins. Neuron 19:939-945.]

Tau and Synuclein Staining:
Here is a general procedure for the immunohistochemical staining of tau, its phosphorylated forms, and synuclein in frozen sections of rat brain. This protocol is adapted from one kindly contributed by Dr. Emil Adamec, M.D., Ph.D., McLean Hospital, Belmont, MA. The procedure was developed for use with cryostat sections of rat brain.
- Sections were fixed with 4% paraformaldehyde for 25 minutes. Sections were then extracted with 0.01% (v/v) Triton X-100. Endogenous peroxidase was blocked with 3% hydrogen peroxide. Sections were then treated with 80% formic acid for 5 to 10 minutes at room temperature to enhance staining. Note that formic acid treatment is also very useful prior to staining with anti-synuclein antibodies.
- The various antibodies were used at dilutions of 1:100 to 1:250. The sections were incubated with the diluted primary antibody overnight at 4°C. A species-specific secondary antibody labeled with HRP was then used with either DAB or AEC to stain the antigens.
- Applicable rabbit and mouse anti-tau and phospho-tau antibodies are: AHB0042, AHB0061, 44-738G, 39-1800, 13-6400, 13-1400, and 18-7461. Anti-synuclein antibodies useful for IHC are: 18-0215, 18-7461, 32-8100, 32-8200, 32-8500, 35-8300, 35-8400, 39-1800, and AHB0261.