The RayBiotech Phospho-Tau (Ser202/Thr205) ELISA is a phosphorylation assay kit for the quantitative measurement of phosphorylated Tau at Ser202/Thr205 in Human, Mouse and Rat serum, plasma and CSF.
RayBio® Phospho-Tau (Ser202/Thr205) Quantitative ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in Human, Mouse & Rat serum, plasma and CSF. By determining phosphorylated Tau protein in your experimental model system, you can verify pathway activation in your sample. You can simultaneously measure numerous different samples without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human, Mouse & Rat phospho-Tau. 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 (Ser202/Thr205) 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 (Ser202/Thr205) 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
Pan Tau Microplate: 96 wells (12 strips x 8 wells) coated with anti-pan- Tau.
Wash Buffer
Phospho Tau Standard Protein
Phospho Detection Antibody Tau (Ser202/Thr205)
HRP-Streptavidin concentrate
Assay Diluent
TMB One-Step Substrate
Stop Solution
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, samples and standards as instructed.
Add 100 μl of each standard and sample to each well. Incubate 2.5 hours at room temperature or overnight at 4ºC with gentle shaking.
Add 100 μl prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
Add 100 μl prepared HRP-Conjugated solution. Incubate for 1 hour at room temperature with gentle shaking.
Add 100 μl TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
Add 100 µl of prepared 1X HRP-Streptavidin to each well.
Add 50 μl Stop Solution to each well. Read at 450 nm immediately.
Typical Data
Standard Curve
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. For prepared reagent storage, see table in manual.
