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User Guide: Clean-Blot IP Detection Kit (HRP)

Version: MAN0011620 17Oct2015
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User Guide: QuantaRed Enhan Chemifluorescent HRP Substrate

Version: Jan. 2015
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Version: Jan. 2015
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Version: Jan. 2015
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Version: Jan. 2015
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Product FAQ

What is the QuantaRed Enhanced Chemifluorescent HRP Substrate?

Answer

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.

Answer Id: E8452

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Product FAQ

What are the Invitrogen AP Chromogenic Substrate (BCIP/NBT) and Invitrogen HRP Chromogenic Substrate (TMB)?

Answer

- Invitrogen AP Chromogenic Substrate (BCIP/NBT) and Invitrogen HRP Chromogenic Substrate (TMB) are ready-to-use, single component substrates for sensitive immunodetection of alkaline phosphatase (AP) or horse radish peroxidase (HRP), respectively, on western blots or dot blots. The sensitivity of the AP substrate is in the 100 pg range, while the HRP substrate is in the 1 ng range (see Note below).

- Invitrogen AP Chromogenic Substrate consists of a ready-to-use solution of 5-bromo-4-chloro-3-indolyl-1-phosphate (BCIP) and nitroblue tetrazolium (NBT) which forms a black-purple precipitate upon reaction with alkaline phosphatase. It is available as a standalone product (Cat. Nos. WP20001) and is also a component of the WesternBreeze Chromogenic Detection Kits.

- Invitrogen HRP Chromogenic Substrate consists of a ready-to-use non-toxic solution of 3,3',5,5'-tetramethylbenzidine (TMB) which forms a blue precipitate upon reaction with horse radish peroxidase. It is available as a standalone product (Cat. No. WP20004).

Note: Sensitivity may vary depending upon the antigen and antibody used for detection.

Answer Id: E10833

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Citations & References

Introduction of a novel HRP substrate-Nanogold probe for signal amplification in immunocytochemistry.

  • Authors: Mayer G, Leone RD, Hainfeld JF, Bendayan M
  • Journal: J Histochem Cytochem (2000) 48:461-470
  • PubMed ID: 10727287
Catalog #
  • L24919
  • N24833(Blocked)
  • T24831(Blocked)

Manual / Product Insert

Diaminobenzidine Histochemistry Kits

Version: 06-30-2004
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Brochure: in vivo diagnostic reagent solution (Chinese)