Tau ELISA Assay Kit
The Tau ELISA Assay Kit is For Research Use Only
Size: 1×96 wells
Sensitivity: 5.2 pg/ml
Dynamic Range: 31.25-2000 pg/ml
Incubation Time: 3 hours
Sample Type: Cell culture supernatant/extracts, serum, plasma, cerebral spinal fluid
Sample Size: 50 µL
Alternative Names: Microtubule-associated Protein Tau, MAPT
This assay employs the quantitative sandwich enzyme immunoassay technique for the quantification of human Tau protein in cell culture supernatant, serum, cell culture extracts, plasma, cerebral spinal fluid. Standards or samples are pipetted into the wells and then add the tagged anti-Tau antibody and HRP-conjugated anti-Tau antibody mixture. Human tau protein will bind to the tagged anti-Tau antibody and HRP-conjugated anti-Tau antibody. The antibodies and Tau protein complex will then bind to the anti-tag antibody coated on the well. After washing away any unbound materials, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of human Tau protein bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450nm ±2nm. The concentration of human Tau protein in the sample is then determined by comparing the O.D of samples to the standard curve.
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This gene encodes the microtubule-associated protein tau (MAPT) whose transcript undergoes complex, regulated alternative splicing, giving rise to several mRNA species. MAPT transcripts are differently expressed in the nervous system, depending on stage of neuronal maturation and neuron type. MAPT gene mutations have been associated with several neurodegenerative disorders such as Alzheimer’s disease, Pick’s disease, frontotemporal dementia, cortico-basal degeneration, and progressive supranuclear palsy.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
- Add 50 μl of standards and samples in duplicate into wells. (Standards and samples should add before antibody mixture)
- Add 50 μl of the Antibody mixture into wells.
- Cover with plate sealer and incubate the plate for 1 hour at room temperature on a plate shaker set to 400 rpm.
- Aspirate each well and wash, repeating the process 2 times for a total 3 washes. Wash by filling each well with 350 μl of 1X Wash Buffer using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels. Take care that none of the wells dry out before the next reagent is dispensed.
- Add 100 μl of TMB substrate subsequently to each well. Incubate for 10 minutes at room temperature in dark on a plate shaker set to 400 rpm.
- Add 100 μl of Stop Solution to each well. Shake plate on a plate shaker for 1 minute to mix.
- Read the OD with a microplate reader at 450 nm immediately.
Typical Standard Curve