Introduction
The RayBio® SUMOylated-SOX9 and Total-SOX9 ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human sumoylated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan SOX9 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and SOX9 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-sumoylation antibody is used to detect sumoylated SOX9 or biotinylated anti-SOX9 antibody is used to detect total SOX9 . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to 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 sumoylated SOX9 or total SOX9 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Product Features
- Quantify quickly and easily
- Screen numerous different lysates without performing a Western Blot analysis
- Minimal hands-on time, convenient, and non-radioactive material
Kit Components
| Component | Size / Qty |
|---|
| Microplate Coated With Anti-Pan-SOX9 Antibody | 1 plate |
| Anti-SUMOylation Detection Antibody | 1 vial |
| Anti-SOX9 Detection Antibody | 1 vial |
| HRP-Conjugated Streptavidin | 200 µL |
| Wash Buffer | 25 mL |
| Assay Diluent | 15 mL |
| Lysis Buffer | 5 mL |
| TMB One-Step Substrate Reagent | 12 mL |
| Stop Solution | 8 mL |
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 sample dilutions
- Distilled or deionized water
- Shaker
Assay Procedure Summary
- Prepare all reagents and samples as instructed.
- Add 100 µL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4°C 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 Streptavidin 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 50 µL 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.
Customization
This assay can be developed for other species (mouse, rat, etc.) upon request. Contact
techsupport@raybiotech.com for more information.
