3,3prime-Diaminobenzidine tetrahydrochloride A widely used chromogenic peroxidase substrate for staining

3,3′-Diaminobenzidine tetrahydrochloride (CAS 868272-85-9)

3,3′-Diaminobenzidine tetrahydrochloride, CAS 868272-85-9 is rated 3.0 out of 5 by 2.
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| See product citations (23)
Synonym: 3,3',4,4'-Tetraaminobiphenyl tetrahydrochloride
Application: A widely used chromogenic peroxidase substrate for staining
CAS Number: 868272-85-9
Molecular Weight: 360.11
Molecular Formula: C12H14N44HCl
* Refer to Certificate of Analysis for lot specific data (including water content).
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Ordering Information

3,3'-Diaminobenzidine tetrahydrochloride sc-209686 1 g $32.00
3,3'-Diaminobenzidine tetrahydrochloride sc-209686A 5 g $104.00

3,3'-Diaminobenzidine tetrahydrochloride hydrate is a water-soluble preparation of 3,3'-Diaminobenzidine, a tetraamine biphenyl compound used as a histological stain for peroxidase activity. Reaction between 3,3'-Diaminobenzidine and peroxidase forms a visible insoluble product. This approach has been used to assay activity of glutamate decarboxylase in rat cerebellar sections and to detect horseradish peroxidase presence on cultured cell surfaces.


1. Saito, K., et al. 1974. Proc. Natl. Acad. Sci. U.S.A. 71: 269-273. PMID: 4131274
2. Pelliniemi, L.J., et al. 1980. J. Histochem. Cytochem. 28: 191-192. PMID: 6986432
3. Messing, A., et al. 1985. J. Histochem. Cytochem. 33: 837-839. PMID: 3894500
4. de Jong, D., et al. 1994. Cancer Res. 54: 256-260. PMID: 8261449

Appearance :
Physical State :
Solubility :
Soluble in water.
Storage :
Desiccate at room temperature
Melting Point :
280° C
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.
WGK Germany :
PubChem CID :
MDL Number :
EC Number :

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3,3prime-Diaminobenzidine tetrahydrochlorideProduct Citations

See how others have used 3,3prime-Diaminobenzidine tetrahydrochloride. Click on the entry to view the PubMed entry .

Citations 1 to 10 of 23 total

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Wang, Z. et al. 2016. Platelet-rich plasma gel composited with nondegradable porous polyurethane scaffolds as a potential auricular cartilage alternative. Journal of biomaterials applications. 30: 889-99.

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Ba, R. et al. 2015. Cell-bricks based injectable niche guided persistent ectopic chondrogenesis of bone marrow-derived mesenchymal stem cells and enabled nasal augmentation. Stem Cell Res Ther. 6: 16.

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Cai, Z. et al. 2015. Chondrogenesis of Human Adipose-Derived Stem Cells by In Vivo Co-graft with Auricular Chondrocytes from Microtia. Aesthetic plastic surgery. 39: 431-9.

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Xie, A. et al. 2014. The application of autologous platelet-rich plasma gel in cartilage regeneration. Mol Med Rep. 10: 1642-8.

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Zhang, L. et al. 2014. Regeneration of human-ear-shaped cartilage by co-culturing human microtia chondrocytes with BMSCs. Biomaterials. 35: 4878-87.

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Chen, JY. et al. 2014. Hepatitis B spliced protein (HBSP) promotes the carcinogenic effects of benzo [alpha] pyrene by interacting with microsomal epoxide hydrolase and enhancing its hydrolysis activity. BMC Cancer. 14: 282.

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Zhao, WJ. et al. 2013. NGX6 expression improves the sensitivity of tamoxifen-resistant MCF-7 cells through modulation of the Smad signaling pathway. Int. J. Oncol.. 42: 2060-8.

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Chen, B. et al. 2013. Macromolecular crowding effect on cartilaginous matrix production: a comparison of two-dimensional and three-dimensional models. Tissue Eng Part C Methods. 19: 586-95.

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Wu, J. et al. 2013. Improvement of PHBV scaffolds with bioglass for cartilage tissue engineering. PLoS ONE. 8: e71563.

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Wang, B. et al. 2013. Proteomic characterization of the late and persistent effects of cadmium at low doses on the rat liver. J Appl Toxicol. 33: 546-57.

Citations 1 to 10 of 23 total
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Rated 1 out of 5 by from Not the IHC substrate either? I wanted the IHC horse radish peroxidase substrate; originally erred in buying diaminobenzidine alone rather than the tetrahydrochloride version. But this tetrahydrochloride will not dissolve in buffer. It does dissolve in water at 0.5mg/ml but if I use or add PBS or TBS, it becomes a cloudy precipitate (I routinely have used the "real" IHC substrate in PBS at 0.5mg/ml and added H2O2 to 0.03%). Even if i filter and use the supernatant (and add H2O2 to 0.03%), it hardly reacts at all when i add pure HRP to it! I have not had this issue with another manufacturer's product (Sigma; was on backorder so i tried Santa Cruz), though that was the "tetrahydrochloride hydrate" CAS 7411-49-6 that i got from Sigma; this Santa Cruz product has a different CAS number 868272-85-9 and I can't get it to work/ dissolve. Therefore, I have to wonder how it's being used by those who buy it because it's NOT for IHC or peroxidase-related reactions on tissue sections.
Date published: 2018-04-11
Rated 5 out of 5 by from Lukenda A; et al Lukenda A; et al., PubMed ID: 26290260 utilized 3,3'-Diaminobenzidine tetrahydrochloride as a chromogen to detect the expression of cancer stem cell markers CD117 (c-kit), CD34, CD20 and CD15. -SCBT Publication Review
Date published: 2015-03-28
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