ELISA Substrate: TMB for HRP

ELISA REAGENTS

ELISA Substrate: TMB for HRP

High-sensitivity, ready-to-use TMB substrate for ELISA assays using horseradish peroxidase (HRP) enzyme conjugates. Designed for colorimetric detection and absorbance measurement at 450 nm.

CAT: 0930-3652-F10 Size: 120 mL Ready-to-Use Solution
ELISA Substrate TMB for HRP, 120 mL, catalog 0930-3652-F10
Product reference: 0930-3652-F10

Product Overview

ELISA Substrate TMB for HRP is a high-sensitivity chromogenic substrate supplied as a ready-to-use solution. It is intended for ELISA applications involving horseradish peroxidase (HRP) enzyme conjugates, including Streptavidin-HRP detection systems.

During the enzymatic reaction, the substrate develops a soluble blue color. Following the addition of an acidic stop solution, the color changes to yellow, allowing absorbance to be measured at 450 nm.

Intended application

Colorimetric detection in ELISA assays using HRP enzyme conjugates.

PACK SIZE
120 mL
DETECTION
450 nm
STORAGE LIFE
At least 12 months*
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*Shelf life: at least 12 months from the date of receipt, according to the supplied product information.

Key Features & Benefits

High-Sensitivity Detection

Supports colorimetric signal development in HRP-based ELISA assays.

Ready-to-Use Format

Supplied as a prepared solution to simplify assay preparation.

Clear Color Transition

Develops blue before stopping and yellow after the addition of acid.

Standard Readout

The stopped reaction is measured at an absorbance wavelength of 450 nm.

Principle of TMB-Based ELISA Detection

In an HRP-based ELISA, horseradish peroxidase catalyzes the oxidation of TMB, producing a blue-colored reaction product. Adding an acidic stop solution terminates the enzymatic reaction and converts the color to yellow. The absorbance of the stopped reaction is then measured at 450 nm.

1. Add TMB

Add substrate after HRP incubation and washing.

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2. Develop Color

Allow the enzymatic reaction to produce a blue color.

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3. Stop & Read

Add acid to turn the solution yellow and read at 450 nm.

General Usage Recommendations

Follow the validated protocol for your ELISA assay. The following recommendations are based on the supplied product information.

  1. Preferably use the substrate cold, in accordance with the assay protocol.
  2. Add the ready-to-use TMB substrate after incubation with HRP and subsequent washing steps.
  3. Allow color development for approximately 5–15 minutes, monitoring the reaction as required by the assay protocol.
  4. Stop the enzymatic reaction using a suitable acid stop solution. The supplied information gives 0.18 M sulfuric acid (H₂SO₄) as an example.
  5. Measure the absorbance of the stopped reaction at 450 nm.
Important: Incubation time, reagent volumes, temperature and stopping conditions should be confirmed against the validated protocol for your specific assay. Handle acidic stop solutions according to the applicable laboratory safety procedures.

Technical Specifications

Product Name ELISA Substrate: TMB for HRP
Catalog Number 0930-3652-F10
Package Size 120 mL
Substrate Type TMB (3,3′,5,5′-tetramethylbenzidine)
Enzyme System Horseradish peroxidase (HRP), including Streptavidin-HRP conjugates
Color Before Stopping Blue
Color After Stopping Yellow
Measurement Wavelength 450 nm
Typical Development Time 5–15 minutes, subject to assay protocol
CAS Number Supplied 7732-18-5
Shelf Life At least 12 months from date of receipt
Dry Ice Required No
Hazardous Shipping Classification No, according to the supplied product listing. Follow the product SDS for laboratory handling.

Note: CAS 7732-18-5 corresponds to water. Confirm the listed CAS number against the manufacturer's documentation before using it as an identifier for the complete formulation.

Research Use Only

This product is intended for research use only and is not for use in diagnostic procedures. Use in accordance with the applicable product documentation and laboratory safety requirements.

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Availability: 24/48H for stock items; 2–6 weeks for non-stock items.

2nd Oct 2026

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