Human MAPtau (Microtubule Associated Protein Tau/Tau Protein) ELISA Kit (HUES02072)

SKU:
HUES02072
Product type:
ELISA
Reactivity:
Human
ELISA Type:
Sandwich
€299

Description

ELISA Kit Technical ManualMSDS

Human MAPtau (Microtubule Associated Protein Tau/Tau Protein) ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human MAPtau . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human MAPtau and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human MAPtau, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm 2 nm. The OD value is proportional to the concentration of Human MAPtau. The concentration of Human MAPtau in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection MethodColormetric
Detection Range7.81-500 pg/mL
Sensitivity4.69 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Human MAPtau in samples. No significant cross-reactivity or interference between Human MAPtau and analogues was observed.

Typical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(pg/mL)
O.D Average Corrected
5002.347
2.403
2.3752.293
2501.685
1.717
1.7011.619
1251.01
0.97
0.990.908
62.50.481
0.503
0.4920.41
31.250.264
0.256
0.260.178
15.630.188
0.178
0.1830.101
7.810.132
0.136
0.1340.052
00.074
0.09
0.082--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human MAPtau were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human MAPtau were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)23.8052.53178.7124.8856.27182.54
Standard deviation1.592.257.401.532.975.57
C V (%)6.684.284.146.155.283.05

Recovery

The recovery of Human MAPtau spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)85-9791
EDTA plasma (n=5)94-10899
Cell culture media (n=5)90-10195

Linearity

Samples were spiked with high concentrations of Human MAPtau and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)90-10590-10290-105
Average (%)969796
1:4Range (%)93-10687-10282-92
Average (%)989387
1:8Range (%)90-10482-9380-94
Average (%)988787
1:16Range (%)87-10381-9083-95
Average (%)948689

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human MAPtau (Microtubule Associated Protein Tau/Tau Protein) ELISA Kit (HUES02072) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100 µL of standard solutions into the standard wells.
    3. Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100 µL of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100 µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37 °C.
    7. Aspirate or decant the solution from the plate and add 350 µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 90 µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37 °C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Human MAPtau (Microtubule Associated Protein Tau/Tau Protein) ELISA Kit (HUES02072) Protein Information

UniProt Protein Function:Tau: a microtubule-associated protein that regulates microtubule assembly and stability. Apparently involved in the establishment and maintenance of neuronal polarity. Mutations can result in several neurodegenerative disorders such as Alzheimer's disease, Pick's disease, frontotemporal dementia, cortico-basal degeneration and progressive supranuclear palsy. The C-terminus binds axonal microtubules while the N-terminus binds neural plasma membrane components, suggesting that tau functions as a linker. Axonal polarity is predetermined by tau localization (in the neuronal cell) in the domain of the cell body defined by the centrosome. Nine differentially spliced isoforms have been described. The short isoforms allow plasticity of the cytoskeleton, whereas the longer isoforms may preferentially play a role in its stabilization.
UniProt Protein Details:

Protein type:Cytoskeletal

Chromosomal Location of Human Ortholog: 17q21.1

Cellular Component: microtubule; microtubule associated complex; growth cone; axon; tubulin complex; plasma membrane; axoneme; cytosol

Molecular Function:protein binding; enzyme binding; structural constituent of cytoskeleton; microtubule binding; apolipoprotein binding; protein kinase binding; SH3 domain binding

Biological Process: axon extension; apoptosis; positive regulation of microtubule polymerization; positive regulation of axon extension; axon cargo transport; neuron migration; microtubule cytoskeleton organization and biogenesis; adult walking behavior; regulation of microtubule polymerization; mitochondrion transport along microtubule; negative regulation of intracellular transport; generation of neurons; regulation of autophagy; cell structure disassembly during apoptosis

Disease: Supranuclear Palsy, Progressive, 1; Pick Disease Of Brain; Frontotemporal Dementia; Parkinson-dementia Syndrome; Parkinson Disease, Late-onset; Frontotemporal Lobar Degeneration With Tdp43 Inclusions, Grn-related

NCBI Summary: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 differentially 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. [provided by RefSeq, Jul 2008]
UniProt Code:P10636
NCBI GenInfo Identifier:6754638
NCBI Gene ID:4137
NCBI Accession:NP_005901
UniProt Related Accession:P10636
Molecular Weight:46kDa
NCBI Full Name:microtubule-associated protein tau isoform 2
NCBI Synonym Full Names:microtubule associated protein tau
NCBI Official Symbol:MAPT
NCBI Official Synonym Symbols:TAU; MSTD; PPND; DDPAC; MAPTL; MTBT1; MTBT2; FTDP-17; PPP1R103
NCBI Protein Information:microtubule-associated protein tau
UniProt Protein Name:Microtubule-associated protein tau
UniProt Synonym Protein Names:Neurofibrillary tangle protein; Paired helical filament-tau
Protein Family:Tautomerase
UniProt Gene Name:MAPT
UniProt Entry Name:TAU_HUMAN
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