3,3-Diaminobenzidine (DAB) tetrahydrochloride
(HB0687)
Product overview
| Name | 3,3-Diaminobenzidine (DAB) tetrahydrochloride |
| Alternative names | DAB, DAB substrate |
| Biological description | Overview DAB tetrahydrochloride is a water soluble form of DAB which is a derivative of benzene and is widely used in the staining of nucleic acids and proteins, particularly in IHC and HC procedures. DAB is oxidized by hydrogen peroxide in the presence of peroxidases such as HRP. Once oxidized, DAB forms a dark brown insoluble precipitate at the reaction site to allow visualisation of the target. This precipitate can be chelated with osmium tetroxide and the color produced can be enhanced by the addition of metals (e.g. nickel, copper, silver and cobalt). Uses & applications In IHC, DAB acts as a substrate for peroxidase enzymes conjugated to a primary or secondary antibody e.g. HRP-conjugated antibodies. DAB is also used for peroxisome and mitochondrial COX activity staining and used to detect the presence and distribution of hydrogen peroxide in plants cells. DAB also enhances staining of iron. |
| Purity | >98% |
| Description | Chromogen staining agent widely used in IHC |
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Images
Figure 1. Parvalbumin positive neurons visualized in rat cerebellum using DAB staining
Figure 2. Tyrosine hydroxylase visualized in mouse striatum using DAB staining
Figure 3. Parvalbumin positive neurons visualized in rat cerebellum using DAB staining
Figure 4. Tyrosine hydroxylase visualized in mouse striatum using DAB staining
Paraffin embedded mouse brain section stained for tyrosine hydroxylase using HB6605 showing the dense network of dopaminergic neurons in the mouse striatum. DAB (HB0687) was used to develop the reaction and Modified Mayer's Hematoxylin (HB6189) was used as a nuclear counterstain.
Figure 1. Parvalbumin positive neurons visualized in rat cerebellum using DAB staining
Figure 2. Tyrosine hydroxylase visualized in mouse striatum using DAB staining
Figure 3. Parvalbumin positive neurons visualized in rat cerebellum using DAB staining
Figure 4. Tyrosine hydroxylase visualized in mouse striatum using DAB staining
Paraffin embedded mouse brain section stained for tyrosine hydroxylase using HB6605 showing the dense network of dopaminergic neurons in the mouse striatum. DAB (HB0687) was used to develop the reaction and Modified Mayer's Hematoxylin (HB6189) was used as a nuclear counterstain.
Biological Data
| Application notes | DAB Staining using HRP Conjugated Secondary Antibodies  Stock solutions:
Working solution:
 Protocol:
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Solubility & Handling
| Storage instructions | -20°C |
| Solubility overview | Soluble in water (2% by mass) |
| Important | This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use |
Chemical Data
| Purity | >98% |
| Chemical name | 3,3'-Diaminobenzidine tetrahydrochloride |
| Molecular Weight | 360.11 |
| Chemical structure | ![3,3-Diaminobenzidine tetrahydrochloride | [868272-85-9] Chemical Structure](https://hellobio.com/media/catalog/product//h/b/hb0687_2.png "3,3-Diaminobenzidine tetrahydrochloride | [868272-85-9]") |
| Molecular Formula | C12H14N4.4HCl |
| CAS Number | 868272-85-9 |
| PubChem identifier | 16211746 |
| SMILES | C1=CC(=C(C=C1C2=CC(=C(C=C2)N)N)N)N.O.Cl.Cl.Cl.Cl |
| InChiKey | DXWSCXIZIHILNP-UHFFFAOYSA-N |
| MDL number | MFCD08273058 |
| Appearance | Off-white to brown solid |
References for 3,3-Diaminobenzidine (DAB) tetrahydrochloride
References are publications that support the biological activity of the product
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3,3'-Diaminobenzidine staining interferes with PCR-based DNA analysis.
Doelle et al (2018) Sci Rep. 8(1) : 1272 -
Detection of Hydrogen Peroxide by DAB Staining in Arabidopsis Leaves.
Daudi et al (2012) Bio Protoc 2(18) : e263 -
Chromogen-based immunohistochemical method for elucidation of the coexpression of two antigens using antibodies from the same species.
Nakata et al (2012) J Histochem Cytochem 60(8) : 611-9 -
Comparison of histological techniques to visualize iron in paraffin-embedded brain tissue of patients with Alzheimer's disease.
van Duijin et al (201 3) J Histochem Cytochem 61(11) : 785-92 -
Comparison of histological techniques to visualize iron in paraffin-embedded brain tissue of patients with Alzheimer's disease.
van Duijin et al (201 3) J Histochem Cytochem 61(11) : 785-92
Publications
These publications cite the use of 3,3-Diaminobenzidine (DAB) tetrahydrochloride purchased from Hello Bio:
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β Bursting in the Retrosplenial Cortex Is a Neurophysiological Correlate of Environmental Novelty Which Is Disrupted in a Mouse Model of Alzheimer's Disease.
Walsh C et al (2022) The Journal of neuroscience : the official journal of the Society for Neuroscience 42 : 7094-7109PubMedID: 35927034
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