The RayBiotech Phospho-TAU (Thr217) Quantitative ELISA Kit is a convenient and sensitive assay to detect phosphorylated Tau at Threonine-217 (p-Tau217) in human cell lysate samples.
Phospho-TAU (Thr217) Quantitative ELISA Kit
Phospho-TAU (Thr217) Quantitative ELISA Kit
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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Species | Human, Mouse, Rat |
| Sensitivity | 0.09 ng/mL |
| Detection Range | 0.37 ng/mL - 90 ng/mL |
| Specificity | This ELISA antibody pair detects human, mouse and rat Phospho TAU (Thr217). |
| Compatible Sample Types | Tissue Lysates, Cell Lysates |
| Accession Number | P10636 |
| Gene Id | 4137 |
| Gene Symbols | MAPT MAPTL MTBT1 TAU |
| Protein Name / Synonyms | Microtubule-associated protein tau (Neurofibrillary tangle protein) (Paired helical filament-tau) (PHF-tau) |
| Quantitative/Semi-Quantitative | Quantitative |
| Solid Support | 96-well Microplate |
| Design Principle | Sandwich-based |
| Method Of Detection | Colorimetric |
| Research Area | Autophagy |
| Shipping Type | Blue ice |
| Storage | -20°C |
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Introduction
RayBio® Phospho-TAU (Thr217) Quantitative ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human, Mouse and Rat phosphorylated TAU (Thr217). An anti-pan TAU antibody has been coated onto a 96-well plate. Standard and Samples are pipetted into the wells and TAU present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-phospho-TAU (Thr217) antibody is used to detect phosphorylated TAU. After washing away unbound antibody, HRP-conjugated streptavidin is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of TAU (Thr217) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Product Features
- Rapidly measure phosphorylated protein in lysates
- Screen numerous different cell lysates without performing a Western Blot analysis
- Minimal hands-on time, convenient, and non-radioactive material
Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Anti-Phospho Antibody
- HRP-Conjugated Secondary Antibody
- Assay Diluent
- TMB One-Step Substrate
- Stop Solution
- Lysis Buffer
- Positive Control Sample
Other Materials Required
- Distilled or deionized water
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare sample dilutions
- Protease and Phosphatase inhibitors
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- Benchtop rocker or shaker
- Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline
- Prepare all reagents and samples as instructed in the manual.
- Add 100 µl of sample or positive control to each well.
- Incubate 2.5 h at RT or O/N at 4 °C.
- Add 100 µl of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared 1X HRP-Streptavidin to each well.
- Incubate 1 h at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
Storage/Stability
Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate, Assay Diluent, TMB One-Step Substrate Reagent, HRP-Streptavidin, Stop Solution and Cell Lysate Buffer should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control should be stored at -70 °C.
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