missing translation for 'onlineSavingsMsg'
Learn More
  1. Home
  2. Products
  3. Immunoassay Reagents and Kits
  4. ELISA Substrates and Reagents
  5. PI15159

Thermo Scientific™ QuantaRed™ Enhanced Chemifluorescent HRP Substrate Kit

Catalog No. PI15159
Change view
Click to view available options
Quantity:
110 ml
1 product options available for selection
Product selection table with 1 available options. Use arrow keys to navigate and Enter or Space to select.
Catalog No. Quantity
PI15159 110 ml
Use arrow keys to navigate between rows. Press Enter or Space to select a product option. 1 options available.
1 options
Catalog No. PI15159 Supplier Thermo Scientific™ Supplier No. 15159
Only null left

Description

Enable sensitivity comparable to enhanced chemiluminescence (ECL) using the most sensitive fluorescent ELISA substrate for peroxidase (HRP).

Thermo Scientific™ QuantaRed Enhanced Chemifluorescent HRP Substrate is the most sensitive fluorescent ELISA substrate, providing sensitivity comparable to enhanced chemiluminescence.

QuantaRed Enhanced Chemifluorescent HRP Substrate enables high-yield and sensitive fluorescent and colorimetric detection of peroxidase activity in the ELISA format. QuantaRed Substrate uses a proprietary technology to increase fluorescent yield and sensitivity of the well-documented ADHP (10-acetyl-3,7-dihydroxyphenoxazine) chemifluorescence reaction. ADHP is a non-fluorescent compound that reacts with horseradish peroxidase (HRP) to produce resorufin, a soluble, highly fluorescent reaction product (excitation/emission approx. 570/585nm). The long wavelength emission of the resorufin minimizes interference from the low wavelength (blue and green) autofluorescence, making it an ideal substrate for quantitating target molecules in biological samples. In addition, the colored reactopm product of QuantaRed Substrate can be measured spectrophotmetrically at 570nm.

Highlights:

  • Sensitive – low picogram to femtogram detection

  • Red-shifted – less interference from autofluorescence generated by biological samples

  • Convenient – compatible with existing filter sets, no need to purchase special filter sets

  • Complete – easy to use; no buffers to prepare or reagent powders to dissolve

  • Flexible – effective stop solution result in stable signal for at least 4 hours

  • Versatile – produces strong signal for easy fluorometric or colorimetric measurement

Note:

Equilibrate all reagents to room temperature before use. Temperature can affect the reaction kinetics.

Specifications

Product Type ELISA Substrate
Quantity 110 ml
Detection Method Fluorescence
Product Line QuantaRed
Substrate Properties Chemical Substrate
Substrate Type HRP (Horseradish Peroxidase)
Content And Storage Sufficient For: 960 wells (10 x 96-well plates) at 100 μL working solution per well
• QuantaRed ADHP Concentrate, 1 mL
• QuantaRed Enhancer Solution, 50 mL
• QuantaRed Stable Peroxide Solution, 50 mL
• QuantaRed Stop Solution, 10 mL

Store at 4°C.

Frequently Asked Questions (FAQs)

Can QuantaRed Enhanced Chemifluorescent HRP Substrate be used for Western blotting?.

No. This is a soluble HRP substrate for ELISA and other plate-based applications.
What antibody concentrations are recommended for use with QuantaRed Enhanced Chemifluorescent HRP Substrate?

The recommended concentrations of antibodies and HRP conjugate are listed below:

Capture Antibody: 1-10 µg/mL
Detection Antibody: 0.05-1 µg/mL
HRP Conjugate: 0.1-0.2 µg/mL
Can a stop solution from another source be used with the QuantaRed Enhanced Chemifluorescent HRP Substrate?

No. The QuantaRed Stop Solution included in the kit must be used to effectively stop the reaction.
Can the QuantaRed Enhanced Chemifluorescent HRP Substrate signal be measured in a kinetic mode without stopping the reaction?

To measure in a kinetic mode, simply omit the Stop Solution. At high HRP concentrations and prolonged reaction times, the fluorescent product may self-quench, which is observed as continuous color development without a change in fluorescent intensity. Optimization of the detection antibody or HRP conjugate is required to determine the linear range of a particular assay system.
What types of plates are compatible with the QuantaRed Enhanced Chemifluorescent HRP Substrate?

Black opaque plates typically produce the best results. White opaque plates also can be used, but might produce higher backgrounds. A variety of other plate types, such as gray or opaque plates with transparent bottoms are also useful with this substrate. Use clear plates if the absorbance of the colored product will be measure instead of the fluorescence.
Can the QuantaRed Enhanced Chemifluorescent HRP Substrate be used colorimetrically?

Yes. The color produced by the reaction is dark magenta and can be measured at 570 +/-10 nm on a colorimetric plate reader.
What type of instrumentation and filter sets are needed for quantifying the QuantaRed Enhanced Chemifluorescent HRP Substrate signal?

The QuantaRed Signal can be measured using standard fluorometers equipped with Cy3 Dye filter sets. Quantitation does not require filters that precisely match the excitation/emission maxima; however, a non-overlapping filter set with a bandpass that includes the excitation/emission spectra is required.
Can other wavelengths for excitation and emission be used to quantify HRP using the QuantaRed Enhanced Chemifluorescent HRP Substrate?

Yes. Wavelengths at 530-575 nm for excitation and 585-630 nm for emission can be used for signal detection.
What is included in the QuantaRed Enhanced Chemifluorescent HRP Substrate Kit?

The kit includes QuantaRed ADHP Concentrate, Enhancer Solution, Stable Peroxide Solution and Stop Solution.
What is the QuantaRed Enhanced Chemifluorescent HRP Substrate?

QuantaRed Enhanced Chemifluorescent HRP Substrate is a proprietary formulation that uses chemical enhancers to increase fluorescence yield and sensitivity of 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) chemifluorescence in plate-based (e.g., ELISA) assays. ADHP is a non-fluorescent compound that reacts with horseradish peroxidase (HRP) in the presence of peroxide to produce resorufin, a soluble, highly fluorescent compound with excitation/emission maxima at 570/585 nm. Unlike chemiluminescent substrates, the QuantaRed Enhanced Chemifluorescent HRP Substrate reaction can be stopped with the fluorescent signal remaining stable for several hours. The long wavelength emission spectrum of resorufin helps minimize interference from lower wavelength autofluorescence (blue and green) that exists in many biological samples.

For Research Use Only. Not for use in diagnostic procedures.