TAU [pS396] Phospho-ELISA Kit, Human

Catalog number: KHB7031

Novex™  Related applications: Protein Assays and Analysis

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Description

The Human Tau [pS396] ELISA research-use-only kit is designed to detect and quantify the level of human Tau protein phosphorylated at serine residue 396 in cell culture supernatants and cerebrospinal fluid (CSF) using 96-well plates and a microplate reader. The assay will recognize both natural and recombinant human Tau [pS396].

Performance characteristics
• Sensitivity: <2 pg/mL
• Standard curve range: 15.6–1,000 pg/mL
• Sample types: cell culture supernatants, cerebrospinal fluid (CSF)
• Species cross-reactivity: human
• Sample volume: 50 μL
• Total assay incubation time: 4 hrs

Principle of the method
The Human Tau [pS396] kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA). A monoclonal antibody specific for all forms of human Tau (either phosphorylated or non-phosphorylated) has been coated onto the wells of the microtiter strips provided. Samples, including standards of known human Tau [pS396] content, control specimens, and unknowns, are pipetted into these wells. During the first incubation, the human Tau [pS396] antigen binds to the immobilized (capture) antibody on one site. After washing, a rabbit polyclonal antibody specific for human Tau [pS396] is added.

During the second incubation, this antibody binds to the immobilized human Tau [pS396] captured during the first incubation. After removal of excess second antibody, a horseradish peroxidase–labeled anti-rabbit IgG antibody is added. This binds to the rabbit polyclonal antibody to complete the four-member sandwich. After a third incubation and washing to remove all the excess anti-rabbit IgG-HRP, a substrate solution is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of human Tau [pS396] present in the original specimen.

Background information
Tau is a microtubule-associated protein of considerable importance to neuronal axons of the vertebrate brain. Tau protein is highly soluble and normally attached to axonal microtubules. Tau stabilizes the microtubules and makes them rigid. Tau interacts with actin in the cytoskeleton and neuronal outgrowth, anchors enzymes such as protein kinases and phosphatases, and regulates intracellular vesicle transport. Tau is phosphorylated by numerous serine/threonine kinases, including GSK-3β, PKA, PKC, CDK5, MARK, JNK, p38MAPK and casein kinase II. Tau phosphorylation regulates both normal and pathological functions of this protein. An over-representation of certain phosphorylated residues on Tau have been reported in neurodegenerative disorders including Alzheimer’s disease (AD), cortical basal degeneration (CBD), progressive supranuclear palsy (PSP), Pick’s disease, and certain forms of Parkinson’s disease.

Hyper-phosphorylated Tau including phosphorylation at serine 396 and serine 404 is the major component of paired helical filaments (PHFs), the building block of neurofibrillary lesions in Alzheimer’s disease (AD). Hyper-phosphorylation impairs the microtubule binding function of Tau, resulting in the destabilization of microtubules in AD brains, ultimately leading to neuronal degeneration. Serine 396 and serine 404 are the major phosphorylation sites of Tau in PHFs. Phosphorylation of serine 396 and serine 404 is primarily responsible for the loss of Tau-mediated tubulin polymerization. Circulating Tau is detectable in cerebrospinal fluid (CSF). Levels of Tau and phosphorylated Tau are reportedly increased in Alzheimer’s disease and other neurodegenerative diseases.

Detect low-expressing proteins and use less sample
ELISA kits designed to measure intracellular signaling targets are typically 2–10 times more sensitive than western blotting. The improved sensitivity enables you to detect low-expressing proteins that otherwise may not be distinguishable from background. In addition, the amount of sample needed to run the assay is less than what is needed for western blots.

Each Human Tau [pS396] ELISA kit is validated for sensitivity, specificity, precision, and lot-to-lot consistency. See product insert for more information on validation.

Related links:
Learn more about ELISA kits
Learn more about other immunoassays
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Gene Aliases: DDPAC, FLJ31424, FTDP-17, MAPTL, MGC138549, MSTD, MTBT1, MTBT2, PPND, TAU
Shipping Condition: Wet Ice
Marker: Tau
Gene ID: 4137
Species: Human
Gene Symbol: MAPT
Sensitivity: <2 pg/mL
Target Gene: microtubule-associated protein tau
Label or Dye: HRP (Horseradish Peroxidase)
Product Size: 96 assays
Concentration: 15.6-1000 pg⁄ml
Sample Volume: 50 µl
Incubation Time: 4 hrs
Detection Method: Colorimetric
Research Category: Neurobiology
Sample Type (Specific): Supernatants, Cerebrospinal Fluid, Tissue Homogenates
For Use With (Equipment): Microplate Reader

Contents & storage

Pre-coated plate(s), standard or calibrator, detector antibody, HRP conjugate, diluents, wash buffer, substrate(s), and stop solution. Store kit at 2-8°C. See product manual for detailed contents and storage conditions for maximum stability.

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Manuals & protocols

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