What are the most common ELISA conjugate/substrate combinations and what are their appropriate absorbance wavelengths?

May 28, 2012

ELISA Enzyme and Substrate Systems

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Dr EJ Dell
PhD, Sales Manager Northwest
Enzyme Conjugate label Substrate Reading wavelength (nm) - Kinetic or Nonstopped Reading wavelength (nm) - End Point or Stopped
Horseradish peroxidase (HRP) OPD (o-Phenylenediamine) 450 492
  TMB (3,3′,5,5′-Tetramethylbenzidine) 630 450
  ABTS ((2,2'-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]) 414 414
Alkaline Phosphatase (Alk-phos) pNPP (p-Nitrophenyl Phosphate) 405 405
β-Galactosidase (β-gal) ONPG (o-Nitrophenyl-ß-D-galactopyranosidase) 420 420
Urease Urea bromocresol 590 590

Here you can find a list of BMG LABTECH instruments that can perform absorbance measurements including ELISA.

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