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Details

Stereochemistry ACHIRAL
Molecular Formula C12H14N4
Molecular Weight 214.2664
Optical Activity NONE
Defined Stereocenters 0 / 0
E/Z Centers 0
Charge 0

SHOW SMILES / InChI
Structure of 3,3'-DIAMINOBENZIDINE

SMILES

NC1=CC=C(C=C1N)C2=CC(N)=C(N)C=C2

InChI

InChIKey=HSTOKWSFWGCZMH-UHFFFAOYSA-N
InChI=1S/C12H14N4/c13-9-3-1-7(5-11(9)15)8-2-4-10(14)12(16)6-8/h1-6H,13-16H2

HIDE SMILES / InChI

Molecular Formula C12H14N4
Molecular Weight 214.2664
Charge 0
Count
MOL RATIO 1 MOL RATIO (average)
Stereochemistry ACHIRAL
Additional Stereochemistry No
Defined Stereocenters 0 / 0
E/Z Centers 0
Optical Activity NONE

Description

(1,1'-Biphenyl)-3,3',4,4'-tetramine (3,3'-Diaminobenzidine, DAB) is an organic benzidine derivative, that used as a precursor to polybenzimidazoles. As its water-soluble tetrahydrochloride, (1,1'-Biphenyl)-3,3',4,4'-tetramine has been used in immunohistochemical staining of nucleic acids and proteins. Diaminobenzidine is oxidized by hydrogen peroxide in the presence of hemoglobin to give a dark-brown color. This color change is used to detect fingerprints in blood. The solubility of (1,1'-Biphenyl)-3,3',4,4'-tetramin in water allows for adaptability compared to other detection solutions which use toxic solvents. In research, this reaction is used to stain cells that were prepared with hydrogen peroxidase enzyme, following common immunocytochemistry protocols. Relevant to Alzheimer's disease, Aβ protein amyloid plaques are targeted by a primary antibody, and subsequently by a secondary antibody, which is conjugated to a peroxidase enzyme. This will bind (1,1'-Biphenyl)-3,3',4,4'-tetramine as a substrate and oxidize it, producing an easily observable brown color. Plaques can then be quantified for further evaluation.

Originator

Approval Year

Targets

Primary TargetPharmacologyConditionPotency

Conditions

ConditionModalityTargetsHighest PhaseProduct

PubMed

Sample Use Guides

In Vivo Use Guide
mouse LD50=100 mg/kg
Route of Administration: Intravenous
In Vitro Use Guide
Pea embryos were dissected from developing seeds of pea plants (Pisum sativum) grown in pots in a greenhouse and irrigated with water. Embryos from pea and leaf fragments from A. thaliana and tomato (Lycopersicon esculentum) were vacuum-infiltrated with the fixation solution containing 2% (w/v) paraformaldehyde, 1% (v/v) glutaraldehyde, 1% (w/v) caffeine in 100 mM phosphate buffer (pH 7) for 30 min and incubated for 15 h in the same solution. The fixed samples were washed with 0.1 M phosphate buffer (pH 7.4) three times and dehydrated in successive baths of 50, 70, 90, 95, and 100 ethanol, butanol/ ethanol 1:1 (v/v), and 100% butanol. Then, the tissues were embedded in the Technovit 7100 resin (Heraeus Kulzer) according to the manufacturer’s instructions, and thin sections (5 mm) were made. The sections were deposited on glass slides that were incubated for 45 min in Perls stain solution, except for negative controls. After washing with distillated water, the glass slides were incubated in a methanol solution containing 0.01 M NaN3 and 0.3% (v/v) H2O2 for 1 h and then washed with 0.1 M phosphate buffer (pH 7.4). For the intensification reaction, samples were then incubated between 10 and 30 min in a 0.1 M phosphate buffer (pH 7.4) solution containing 0.025% (w/v) DAB (Sigma), 0.005% (v/v) H2O2, and 0.005% (w/v) CoCl2 (intensification solution). Rinsing with distilled water stopped the reaction
Substance Class Chemical
Record UNII
2RV4T6KHQI
Record Status Validated (UNII)
Record Version