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QuantaBlu Fluorogenic Peroxidase Substrate


A rapid and sensitive fluorescent ELISA substrate for HRP.

QuantaBlu Chemifluorescent ELISA Substrate

Thermo Scientific QuantaBlu Fluorogenic Peroxidase Substrate is a soluble fluorogenic (chemifluorescent) substrate for detection of peroxidase activity in ELISA methods.

QuantaBlu Fluorogenic Peroxidase Substrates allow rapid stopped and kinetic ELISA assays to be performed with greater sensitivity than TMB and OPD substrates. Two formulations are offered, one with slow development for kinetic assays and one optimized for stopped (non-kinetic) assays.

Highlights:

  • More sensitive than TMB, OPD or ABTS substrates
  • Flexible - stopped, non-stopped or kinetic assays possible
  • Large dynamic range (4 log peroxidase concentration range)
  • Excellent stability: working solution is stable for 24 hours
  • Large stokes shift; excitation/emission maxima of 325/420; range of 315-345/370-460
  • Does not photobleach

Product Details:

A variety of substrates are available for detecting peroxidase activity in ELISA-based assays. Colorimetric substrates (e.g., TMB, OPD and ABTS) have been widely used for years. Each of these substrates varies greatly with respect to its performance characteristics such as detection sensitivity, working range and attainable signal-to-noise ratios. Substrate flexibility is also a key issue that affects an assay. Stability, development time requirements and the capacity to perform stopped and/or kinetic assays vary significantly among these substrates. Ideally, a substrate is stable, very sensitive, has high attainable signal-to-noise ratios, possesses a broad dynamic range, and allows the user to perform stopped, non-stopped and kinetic assays.

QuantaBlu Fluorogenic Peroxidase Substrate meets all the requirements of an ideal substrate for use in peroxidase detection. QuantaBlu Substrate generates a blue fluorescent product upon reaction with peroxidase. The fluorescent product does not photobleach. Fluorometric-based detection overcomes the limitations of colorimetric substrate detection, which does not allow for quantitation of greater than four optical density units. QuantaBlu Substrate allows for stopped, non-stopped and kinetic assays to be performed. Incubation times with QuantaBlu Substrate for stopped and non-stopped assays may be varied between one to 90 minutes at either room temperature or 37°C. QuantaBlu Substrate exhibits a flat baseline in assays, which facilitates low-level detection sensitivity and allows for high signal-to-noise ratios.

Comparison of QuantaBlu fluorogenic substrate to TMB ABTS and OPD ELISA Substrates
Comparison of Thermo Scientific QuantaBlu Fluoregenic Peroxidase Substrate to TMB, ABTS and OPD ELISA substrates. QuantaBlu fluorescent substrate and other colorimetric substrates were incubated for 30 minutes at room temperature with increasing amounts of horseradish peroxidase enzyme followed by addition of a stop solution. QuantaBlu Fluorogenic Peroxidase Substrate gave the greatest signal-to-noise ratios and exhibited the lowest detection limit.

Rapid and sensitive detection with QuantaBlu Fluorogenic Peroxidase Substrate for Kinetic ELISA Assays
Thermo Scientific QuantaBlu Fluorogenic Peroxidase Substrate provides the ability to rapidly detect peroxidase at very low concentrations in short periods of time. QuantaBlu fluorescent substrate was read in a non-stopped mode using a 1-minute instrument cycle time between reads. The graph below illustrates the detection of horseradish peroxidase enzyme amounts ranging from 0-10pg per well from 1.5-6.5 minutes of substrate incubation time. At 1.5 minute, as little as 2.5pg of peroxidase was detected. At 6.5 minutes, 0.625pg of peroxidase was detected.

References:

  1. Savage, M.D., et al. (1998). QuantaBlu Fluorogenic Peroxidase Substrate: the ideal peroxidase substrate. Previews 2(1): 6-9.
  2. Savage, M.D., et al. (1998). QuantaBlu Fluorogenic Peroxidase Substrate offers rapid data acquisition and high sensitivity. Previews 2(2): 18-9.
  3. Ayala, P., et al. (2002). Infect. Immun. 70: 5965-71.
  4. Atamna, H., et al. (2000). Proc. Natl. Acad. Sci. USA. 97: 686-91.
  5. Jefcoat, A.M., et al. (2001). Am. J. Physiol. Lung Cell. Mol. Physiol. 281: L704-12.

Related Resources:

Assay development and ELISA technical guide
ELISA substrate selection guide

Related Products:

QuantaRed Enhanced Fluorogenic Peroxidase Substrate


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
15162 QuantaBlu NS/K Fluorogenic Substrate Kit
Formulation: Two-component kit containing proprietary fluorescent substrate and peroxide solutions for non-stopped kinetic assays
Sufficient For: 275mL of 1X working solution (2750 wells at 100µL per well)

Kit Contents:
QuantaBlu Substrate Solution, 250mL
QuantaBlu Peroxide Solution, 30mL
280mL kit Product Instructions for product #15162 QuantaBlu NS/K Fluorogenic Substrate Kit MSDS for product #15162 QuantaBlu NS/K Fluorogenic Substrate Kit Certificate of Analysis for product #15162 QuantaBlu NS/K Fluorogenic Substrate Kit $182.00

Add to cart
15169 QuantaBlu Fluorogenic Peroxidase Substrate Kit
Formulation: Three-component kit containing proprietary fluorescent substrate, peroxide and stop solutions for stopped (endpoint) or non-stopped assays
Sufficient For: 275mL of 1X working solution (2750 wells at 100µL per well)

Kit Contents:
QuantaBlu Substrate Solution, 250mL
QuantaBlu Peroxide Solution, 30mL
QuantaBlu Stop Solution, 275mL
550mL kit Product Instructions for product #15169 QuantaBlu Fluorogenic Peroxidase Substrate Kit MSDS for product #15169 QuantaBlu Fluorogenic Peroxidase Substrate Kit Certificate of Analysis for product #15169 QuantaBlu Fluorogenic Peroxidase Substrate Kit $213.00

Add to cart
15169B QuantaBlu Fluorogenic Peroxidase Substrate Kit
Formulation: Three-component kit containing proprietary fluorescent substrate, peroxide and stop solutions for stopped (endpoint) or non-stopped assays

Kit Contents:
QuantaBlu Substrate Solution
QuantaBlu Peroxide Solution
QuantaBlu Stop Solution
Custom MSDS for product #15169B QuantaBlu Fluorogenic Peroxidase Substrate Kit

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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