The Human Tau (Total) ELISA research-use-only kit is designed to detect and quantify the level of human Tau protein (phosphorylated and nonphosphorylated) 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.
Performance characteristics
• Sensitivity: <12 pg/mL
• Standard curve range: 31.2–2,000 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 (Total) kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA). A monoclonal antibody specific for human Tau has been coated onto the wells of the microtiter strips provided. Samples, including standards of known human Tau content, control specimens, and unknowns, are pipetted into these wells. During the first incubation, the human Tau antigen binds to the immobilized (capture) antibody on one site. After washing, a rabbit polyclonal antibody specific for human Tau is added.
During the second incubation, this antibody binds to the immobilized human Tau captured during the first incubation. After removal of excess second antibody, a horseradish peroxidase-labeled anti-rabbit 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-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 present in the original specimen.
Background information
Tau is a microtubule-associated protein found predominantly in neuronal axons of the vertebrate brain. Human tau exists as six different isoforms that result from alternative splicing of the single transcript derived from a gene located on chromosome 17. The molecular weight of the tau isoforms ranges from 48 kDa to 68 kDa. 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.
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 (Total) ELISA kit is validated for sensitivity, specificity, precision, and lot-to-lot consistency. See protocol insert for more information on validation.
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