Introduction
RayBio® Phospho-TAU (Ser199) Quantitative ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human, Mouse phosphorylated TAU (Ser199). 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 rabbit anti-phospho-TAU (Ser199) antibody is used to detect phosphorylated TAU. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG 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 (Ser199) 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
Kit Components
- Pre-coated 96-well microplate
- Lyophilized Standard
- Anti-phospho Detection Antibody
- HRP-Conjugated Secondary Antibody
- Wash Buffer
- Assay Diluent
- Lysis Buffer
- TMB One-Step Substrate Reagent
- Stop Solution
Additional Materials Required
- Microplate reader capable of measuring absorbance at 450 nm
- Protease and Phosphatase inhibitors
- Precision pipettes to deliver 2 µL to 1 mL volumes
- Adjustable 1-25 mL pipettes for reagent preparation
- 100 mL and 1 liter graduated cylinders
- Log-log graph paper or computer and software for ELISA data analysis
- Absorbent paper
- Tubes to prepare the positive control or sample dilutions
- Distilled or deionized water
- Shaker
Assay Procedure Summary
- Prepare all reagents, samples and positive control as instructed.
- Add 100 ÎĽL of each standard and sample to each well. Incubate 2.5 hours at room temperature or overnight at 4ÂşC with gentle shaking.
- Add 100 ÎĽL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
- Add 100 µL prepared HRP-conjugated secondary antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
- Add 100 ÎĽL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
- Add 50 ÎĽL Stop Solution to each well. Read at 450 nm immediately.
Storage/Stability
The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.
