TMB/H2O2 Substrate for ELISA

Product Characteristics

TMB (3, 3′, 5, 5′-tetramethylbenzidine) is a chromogenic substrate for enzyme horseradish peroxidase
(HRP). During the enzymatic degradation of hydrogen peroxide by HRP, TMB produces a deep blue
color due to formation of a one-electron oxidation product. This compound, a cation free radical, has
an adsorption maxima at 653 nm. Further, acidification of the radical with acid yields the diimine
adsorbing light at 450 nm.

₹900.00 – ₹5,000.00

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Composition & Properties

Ready-to-use substrate: Includes substrate buffer, TMB and hydrogen peroxide.

Working Procedure

  • The desired amount of substrate is poured into a container and allowed to reach room
    temperature in the dark before use.
  • A recommended volume of 100 µl substrate solution per microtiter well is added. Time of
    colour development is typically 5-20 minutes. The plate to be kept in the dark during incubation.
  • Add 100 µl (equal volumes) of 0.50 M sulphuric acid and the absorbance is read at 450/620 nm
    immediately.

Tips & Tricks

  • If reduced intensity is required, it is recommended to shorten the time of development or to
    dilute the conjugate/ antibody in the immunoassay further.
  • Dilution of the substrate is not recommended.

Handling & Storage

  • Store solution at 2-8 °C in the dark.
  • Avoid exposure to light, heat and contamination with metal ions or peroxidase.
  • Re-dispense only into amber colour bottles made of High Density Polyethylene (HDPE).
CERTIFICATE OF ANALYSIS
Quantity

100mL, 250mL, 500mL, 1000mL

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