TMB Substrate Solution for HRP Conjugates
TMB substrate solution is a ready-to-use chromogenic substrate designed for horseradish peroxidase (HRP) conjugated systems. It is widely used in ELISA, immunoassays, and other HRP-based detection methods for sensitive and reproducible signal generation.
Principle of Reaction
HRP catalyzes the oxidation of TMB, producing a soluble blue-colored product. Upon addition of a stop solution, typically sulfuric, the reaction converts to a yellow end product with a measurable absorbance at 450 nm. The intensity of the color is directly proportional to the amount of HRP conjugate present.
Performance Characteristics
Differences in TMB substrate solution formulations affect background and sensitivity of ELISA tests. This TMB substrate solution is formulated for high sensitivity and low background, enabling clear signal differentiation even at low analyte concentrations. The reaction provides:
- Rapid color development
- High signal-to-noise ratio
- Consistent performance across assay runs
Applications
- ELISA (enzyme-linked immunosorbent assay)
- Sandwich and competitive immunoassays
- HRP-based detection systems
- Diagnostic and research laboratory assays
Ready-to-Use Format
Supplied as a stabilized, single-component solution, eliminating the need for mixing or preparation. This reduces variability and improves assay reproducibility.
Handling and Storage
Store at 2–8°C and protect from light. Do not freeze. Allow the solution to reach room temperature before use. Avoid contamination by using clean pipette tips and containers.
Typical Workflow
- Add TMB substrate to wells containing HRP conjugate
- Incubate for appropriate development time (typically 5–20 minutes)
- Add stop solution
- Measure absorbance at 450 nm
Quality and Consistency
Manufactured under controlled conditions to ensure lot-to-lot consistency and reliable assay performance. Formulation includes stabilizers for longer shelf life.