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Substrate Buffers

Substrate buffers for alkaline phosphatase and horseradish peroxidase.

Stable Peroxide Substrate Buffer

The Thermo Scientific Pierce Substrate Buffers include optimized, reliable and convenient stock solutions of peroxide for HRP-based assays and diethanolamine for alkaline phosphatase (alk-phos) detection.


  • Ready-to-use
  • Minimize assay-to-assay variability

Product Details:

Buffered stable hydrogen peroxide (H2O2) is used by researchers who prefer to make their own HRP-ELISA substrates. This is easily done by adding a chromogen of choice to the buffer. Our Stable Peroxide Substrate Buffer has a long shelf life (12 months after receipt) and is provided as a 10-fold concentrate. A total volume of 1000mL can be made from one 100mL bottle of concentrate. If you use 10mL of substrate per plate, this will allow you to make substrate for 100 plates from only one bottle of Stable Peroxide Substrate Buffer.

Diethanolamine Substrate Buffer is used with soluble alkaline phosphatase substrates such as PNPP. Our formulation is convenient, ready-to-use and reduces the possibility of assay-to-assay variability. Diethanolamine Substrate Buffer is sold as 5X concentrate with an optimal, consistent pH. It is stable even at a 1X concentration.


  1. Pifer, J., et al. (2002). J. Immunol. 169: 1372-8.
  2. Neisewander, J.L., et al. (2000). J. Neurosci. 20: 798-805.
  3. Louis, H., et al. (2000). J. Histochem. Cytochem. 48: 499-508.

Related Resources:

Assay development and ELISA technical guide
ELISA substrate selection guide

Ordering Information
Product # Description Pkg. Size Instructions MSDS CofA Price
34062 Stable Peroxide Substrate Buffer (10X)
Formulation: Proprietary buffered solution containing peroxide
Sufficient For: 1L of typical 1X buffer for HRP substrates
100mL Product Instructions for product #34062 Stable Peroxide Substrate Buffer (10X) MSDS for product #34062 Stable Peroxide Substrate Buffer (10X) Certificate of Analysis for product #34062 Stable Peroxide Substrate Buffer (10X) $56.00

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34064 Diethanolamine Substrate Buffer (5X)
Formulation: 5.1M diethanolamine, pH 9.8
Sufficient For: 1.125L typical 1X buffer for AP substrates
225mL Product Instructions for product #34064 Diethanolamine Substrate Buffer (5X) MSDS for product #34064 Diethanolamine Substrate Buffer (5X) Certificate of Analysis for product #34064 Diethanolamine Substrate Buffer (5X) $62.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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