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ReadiUse™ Stayright™ Purple *HRP Chromogen Premixed with Hydrogen Peroxide*
3,3'-Diaminobenzidine (DAB) has been used as the most commonly used IHC chromogen because it is inexpensive and sensitive for routine applications. However, DAB has been shown to be mutagenic and hazardous to laboratory workers and the environment. In order to address this issue, AAT Bioquest has developed this novel Stayright™ Purple as a significantly safer IHC chromogen than DAB. Furthermore, Stayright™ Purple provides a rapid and simple method to develop clean and intense purple stain in the presence of HRP with high sensitivity as DAB. The ready-to-use Stayright™ Purple HRP substrate also shows non-mutagenic effects with minimal cytotoxicity. ReadiUse™ Stayright™ Purple Peroxidase (HRP) Substrate is suitable for use in peroxidase (HRP)-based immunohistochemistry (IHC) and in situ hybridization (ISH) staining methods. The substrate is a stable pre-mixed solution containing hydrogen peroxide so all mixing steps are eliminated and is ready to use. Upon HRP-induced oxidation, Stayright™ Purple forms a purple insoluble precipitating product at the target site of your assay. The purple end product is insoluble in organic solvents and organic mounting media, thus the distinct purple stain can maintain through regular dehydration and coverslipping steps.
Example protocol
AT A GLANCE
Protocol Summary
- Apply ready-to-use Stayright™ Purple solution to tissue section.
- Incubate tissue section for 5-15 minutes.
- Rinse tissue for 5-10 minutes, counterstain.
- Add mounting medium to cover the section.
SAMPLE EXPERIMENTAL PROTOCOL
- Cover section with the ready-to-use Stayright™ Purple solution. Incubate at room temperature for 5-15 minutes.
- Immerse slides in dH2O to stop the color development and monitor the staining intensity. If the staining intensity is not bright enough, longer incubation is needed. One can re-apply the Stayright™ Purple solution to continue the development.
- Wash with dH2O for 5-10 minutes.
- Use a desired counterstain if needed.
- Dehydrate with ethanol and permanently mount in organic permanent mounting medium.
Note For guidelines on sample preparation, please visit https://www.aatbio.com/resources/guides/paraffin-embedded-tissue-immunohistochemistry-protocol.html
Citations
View all 26 citations: Citation Explorer
Epigenetic suppression of SLFN11 in germinal center B-cells during B-cell development
Authors: Moribe, Fumiya and Nishikori, Momoko and Takashima, Tsuyoshi and Taniyama, Daiki and Onishi, Nobuyuki and Arima, Hiroshi and Sasanuma, Hiroyuki and Akagawa, Remi and Elloumi, Fathi and Takeda, Shunichi and others,
Journal: PloS one (2021): e0237554
Authors: Moribe, Fumiya and Nishikori, Momoko and Takashima, Tsuyoshi and Taniyama, Daiki and Onishi, Nobuyuki and Arima, Hiroshi and Sasanuma, Hiroyuki and Akagawa, Remi and Elloumi, Fathi and Takeda, Shunichi and others,
Journal: PloS one (2021): e0237554
3,3'-diaminobenzidine (DAB)-H2O2-HRP voltammetric enzyme-linked immunoassay for the detection of carcionembryonic antigen
Authors: Zhang, S., Yang, J., Lin, J.
Journal: Bioelectrochemistry (2008): 47-52
Authors: Zhang, S., Yang, J., Lin, J.
Journal: Bioelectrochemistry (2008): 47-52
HistoGreen: a new alternative to 3,3'-diaminobenzidine-tetrahydrochloride-dihydrate (DAB) as a peroxidase substrate in immunohistochemistry?
Authors: Thomas, M. A., Lemmer, B.
Journal: Brain Res Brain Res Protoc (2005): 107-18
Authors: Thomas, M. A., Lemmer, B.
Journal: Brain Res Brain Res Protoc (2005): 107-18
Stability and solubility of 3,3'-diaminobenzidine (DAB)
Authors: Kiernan, J. A.
Journal: Biotech Histochem (2003): 135
Authors: Kiernan, J. A.
Journal: Biotech Histochem (2003): 135
Safe diaminobenzidine (DAB) disposal
Authors: Horn, H.
Journal: Biotech Histochem (2002): 229
Authors: Horn, H.
Journal: Biotech Histochem (2002): 229
Page updated on May 19, 2025
Safety data sheet