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3 3 diaminobenzidine tetrahydrochloride dab

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3,3-diaminobenzidine tetrahydrochloride (DAB) is a chromogenic substrate used in immunohistochemistry and other biochemical applications. It produces a brown, insoluble reaction product upon oxidation, which allows for the visualization of target proteins or other analytes in biological samples.

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Lab products found in correlation

Immunohistomount, by Santa Cruz Biotechnology (2 mentions) Hematoxylin, by Thermo Fisher Scientific (1 mentions) Apotag peroxidase in situ apoptosis detection kit, by Merck Group (1 mentions) Caerulein, by Merck Group (1 mentions) Supersignal west femto chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Recombinant dnase 1, by Roche (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Leupeptin, by Roche (1 mentions) Aprotinin, by Roche (1 mentions) Collagenase p, by Roche (1 mentions) Collagenase, by Worthington (1 mentions) Boc gln ala arg amc, by Bachem (1 mentions) Protein determination reagent, by Bio-Rad (1 mentions) Multi screen 96 well plates, by Merck Group (1 mentions) Alkaline phosphatase labeled streptavidin, by BD (1 mentions) Thimerosal, by Merck Group (1 mentions) Ethylenediaminetetraacetic acid edta, by Merck Group (1 mentions) Ferrous sulfate feso4.7h2o, by Thermo Fisher Scientific (1 mentions) Nitrocellulose membrane ff120 hp, by GE Healthcare (1 mentions) Polyvinylpyrrolidone pvp mw 29 000, by Merck Group (1 mentions)

Spelling variants of this product

DAB (142 citations) Diaminobenzidine (DAB) (29 citations) 3,3’-diaminobenzidine tetrahydrochloride (DAB) substrate (5 citations) 3,3′-Diaminobenzidine (DAB; (3 citations) 3,3’-diaminobenzidine tetrahydrochloride plus (DAB+) (3 citations) 3,3′-diaminobenzidine tetrahydrochloride (2 citations) 3,3′-Diaminobenzidine tetrahydrochloride (DAB) immunostaining detection reagent (2 citations)

8 protocols using 3 3 diaminobenzidine tetrahydrochloride dab

1

Immunohistochemical Detection of COX-2

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Tissues were collected in 10% buffered formalin and were embedded in paraffin for preparation as 5-μm tissue sections. All tissue sections were subjected to antigen retrieval in 0.1 mM Citric acid for 8 minutes, and were treated in 10% H2O2 for 10 minutes to block endogenous peroxidase activity.
Immunhistochemical staining was performed using an affinity purified primary rabbit anti-mouse COX-2 antibody (Cayman Chemical, # 160126; 1 μg/ml), which is detected using the ImmPRESS HRP anti Rabbit Ig (Peroxidase) Kit (Vector Laboratories, Burlingame, CA), and visualized with 3,3′ diaminobenzidine tetrahydrochloride (DAB) (Invitrogen, Carlsbad, CA). Sections were counterstained with hematoxylin (Thermo Fisher Scientific, Waltham, MA).
Ra H., González-González E., Uddin M.J., King B.L., Lee A., Ali-Khan I., Marnett L.J., Tang J.Y, & Contag C.H. (2015). Detection of Non-Melanoma Skin Cancer by in vivo Fluorescence Imaging with Fluorocoxib A. Neoplasia (New York, N.Y.), 17(2), 201-207.
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2

Quantification of Apoptosis in Lung Tissue

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Apoptosis in paraffin-embedded lung tissue samples was quantified by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL)77 (link),78 (link). The TUNEL assay was performed on tissue sections as per the manufacturer’s protocol (ApoTag Peroxidase In Situ Apoptosis Detection Kit; Millipore), and the apoptotic cells were detected with 3,3′-diaminobenzidine tetrahydrochloride (DAB; Invitrogen) with hematoxylin counterstaining. For each section, at least six 20 × fields were chosen randomly. The numbers of TUNEL-positive and TUNEL-negative cells were quantified using ImageJ software (NIH) in a blinded manner. The percentage of TUNEL + cells in each section was calculated, and the mean ± SEM for each sample was determined. Tissue sections from a minimum of 5 animals from at least 2 independent experiments were analyzed.
Saunders R.A., Michniacki T.F., Hames C., Moale H.A., Wilke C., Kuo M.E., Nguyen J., Hartlerode A.J., Moore B.B, & Sekiguchi J.M. (2021). Elevated inflammatory responses and targeted therapeutic intervention in a preclinical mouse model of ataxia-telangiectasia lung disease. Scientific Reports, 11, 4268.
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3

Pancreatic acinar cell isolation

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Caerulein (#C9026), trypsin inhibitor (SBTI) (#T9003), protease (#P5147) were purchased from Sigma Aldrich (St Louis, MO), collagenase (CLSPA) was purchased from Worthington Biochemical Co. (Lakewood, NJ), collagenase P (11213857001), DNase I recombinant (04716728001), leupeptin and aprotinin were purchased from Roche Diagnostics (Indianapolis, IN), bovine albumin serum fraction V (BSA) from MP Biomedicals (Solon, OH), Phadebas Amylase Assay kit was purchased form Magle Life Sciences (Cambridge, MA), Boc-Gln-Ala-Arg-AMC from BachemAG (Bubendrof, Switzerland), Dulbecco’s minimal essential medium (DMEM)/high glucose from HyClone (Logan, Utah), TRIzol, and 3,3-diaminobenzidine tetrahydrochloride (DAB) from Thermo Scientific (Rockford, IL). Protein determination reagent was purchased from Bio-Rad Life Science Research (Hercules, CA). Supersignal West Femto Chemiluminescent Substrate was purchased from Thermo Fisher Scientific (Rockford, IL).
Xia D., Halder B., Godoy C., Chakraborty A., Singla B., Thomas E., Shuja J.B., Kashif H., Miller L., Csanyi G, & Sabbatini M.E. (2019). NADPH oxidase 1 mediates caerulein-induced pancreatic fibrosis in chronic pancreatitis. Free radical biology & medicine, 147, 139-149.
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4

Quantifying RSV-specific Antibody and Cytokine Responses

Cited 2 times
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Splenocytes, lung, and bone marrow cells were isolated from tissue samples at 5 d.p.c.. RSV F-specific antibody secreting cells were determined on Multi-screen 96 well plates (Millipore, Billerica, MA) pre-coated with purified recombinant F protein (400ng/ml) by enzyme-linked immunospot (ELISpot) assay as previously described [42 (link)]. The spots of anti-F secreting cells were developed with biotinylated anti-mouse IgG and streptavidin-AP (Southern) and 3,3’-diaminobenzidine tetrahydrochloride (DAB, Thermo Scientific). The spots of IFN-γ and IL-4 secreting cells were determined on Multi-screen 96 well plates coated with cytokine specific capture antibodies as described [40 (link)–42 (link)]. Briefly, lung cells (0.2x106) per well were cultured with peptide F92-106 (ELQLLMQSTPATNNR, 4 μg/ml), F85-93 (KYKNAVTEL), G183-195 (WAICKRIPNKKPG), and M282-90 (SYIGSINNI) as previously described [43 (link), 44 (link)]. After 36 h stimulation, the spots were developed with biotinylated mouse IFN-γ or IL-4 antibody, alkaline phosphatase labeled streptavidin (BD Pharmingen) and then counted using an ELISpot reader (BioSys, Miami, FL).
Kim K.H., Lee Y.T., Hwang H.S., Kwon Y.M., Jung Y.J., Lee Y., Lee J.S., Lee Y.N., Park S, & Kang S.M. (2015). Alum Adjuvant Enhances Protection against Respiratory Syncytial Virus but Exacerbates Pulmonary Inflammation by Modulating Multiple Innate and Adaptive Immune Cells. PLoS ONE, 10(10), e0139916.
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5

Rapid Mycobacterium tuberculosis Detection

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Anti-lipoarabinomannan (LAM) monoclonal capture and detection antibodies were obtained from Chatterjee Lab repository at CSU. Anti-detection antibody was purchased from Sigma Aldrich. LAM was obtained from the Chatterjee Lab repository that was isolated and purified prior from Mycobacterium tuberculosis (Mtb) CDC1551 strain from in vitro culture. Normal urine (NEU) samples were collected from a healthy human volunteer from a TB non-endemic region. Horseradish peroxidase (HRP) conjugation kit-Lightning-Link (ab102890) was purchased from Abcam. 3,3′-diaminobenzidine tetrahydrochloride (DAB) was ordered from Thermo Scientific. Hydrogen peroxide (H2O2), bovine serum albumin (BSA), phosphate buffer saline tablet (PBS), Tween 20, Thimerosal, Ethylenediaminetetraacetic acid (EDTA), and Polyvinylpyrrolidone (PVP; MW 29,000) were obtained from Sigma Aldrich. Triton X-100 and ferrous sulfate (FeSO4•7H2O) were purchased from Fisher Scientific. Trehalose was obtained from Calbiochem. Milli-Q (MQ) water was used to prepare all reagents. Nitrocellulose membrane (FF120HP, GE), absorbent pad (AP30034P0, Millipore), diagnostic microfluidic hydrophilic film (9962, 3M), and adhesive transfer tape (468MP, 3M), which has a great chemical resistance, were used to fabricate the device.
Panraksa Y., Jang I., Carrell C.S., Amin A.G., Chailapakul O., Chatterjee D, & Henry C.S. (2022). Simple manipulation of enzyme-linked immunosorbent assay (ELISA) using an automated microfluidic interface. Analytical methods : advancing methods and applications, 14(18), 1774-1781.
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6

Quantification of Adipose Tissue Hyaluronan

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Epididymal, subcutaneous and interscapular brown AT were removed from mice and fixed for 24 hours at 4°C in Roti®-Histofix 4% (Carl Roth GmbH & Co KG, Karlsruhe, Germany). Subsequently, they were dehydrated and embedded in paraffin. Embedded AT was sliced into tissue sections of a thickness of 5 μm. All lymph nodes were removed prior to analysis. HA was visualized by affinity histochemistry using biotinylated HA-binding protein (Calbiochem, San Diego, CA, USA), detected with a horseradish peroxidase (HRP)-streptavidin conjugate (Sigma-Aldrich, St. Louis, MO, USA) and 3, 3’-diaminobenzidine tetrahydrochloride (DAB) (Thermo Fisher Scientific). Immunostaining was quantified by the ImageJ Software (National Institutes of health, USA) and %area fraction in ROI was normalized to 100 cells.
Misiou A., Garmey J.C., Hensien J.M., Harmon D.B., Osinski V., McSkimming C., Marshall M.A., Fischer J.W., Grandoch M, & McNamara C.A. (2020). Helix-Loop-Helix Factor Id3: A Novel Regulator of Hyaluronan-mediated Adipose Tissue Inflammation. Arteriosclerosis, thrombosis, and vascular biology, 41(2), 796-807.
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7

Immunostaining of Adrenocortical Cytochrome P450 2A6

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Five μm thick FFPE sections of histologically confirmed ACC and normal adrenal tissue were used in this study. Representative ACC (n = 2) and normal adrenocortical samples (n = 2) were selected for immunostaining. Using a standard immunohistochemistry protocol, target epitopes were detected using mouse monoclonal antibody to cytochrome P450 2A6 (Abcam Inc., Cambridge, MA) followed by bovine anti-mouse IgG-HRP conjugated secondary antibody (Santa Cruz, Dallas, TX). DAB (3,3’-diaminobenzidine tetrahydrochloride) was utilized for antigen detection (Life Technologies). Sections were counterstained with hematoxylin (VWR International, Radnor, PA) and mounted using immunohistomount (Santa Cruz). Anonymous samples of histologically normal liver were used as a positive control for CYP2A6 expression.
Murtha T.D., Brown T.C., Rubinstein J.C., Haglund F., Juhlin C.C., Larsson C., Korah R, & Carling T. (2017). Overexpression of Cytochrome P450 2A6 in Adrenocortical Carcinoma. Surgery, 161(6), 1667-1674.
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8

CYP4B1 Immunostaining of Adrenal Tissues

Cited 1 time
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Five μm thick sections of histologically confirmed ACC, ACA, and normal adrenal tissue from formalin fixed paraffin embedded (FFPE) tissue samples were collected for study. Two representative samples of each tissue type were selected for immunostaining. Using standard immunohistochemistry protocols, target epitopes were detected using rabbit anti-CYP4B1 polyclonal antibody (Santa Cruz, Dallas, TX) followed by anti-rabbit HRP conjugated monoclonal secondary antibody (Santa Cruz).22 (link) DAB (3,3’-diaminobenzidine tetrahydrochloride) was utilized for antigen detection (Life Technologies, Carlsbad, CA). Sections were counterstained with hematoxylin and mounted using immunohistomount (Santa Cruz).
Murtha T.D., Korah R, & Carling T. (2016). Suppression of Cytochrome P450 4B1: An Early Event in Adrenocortical Tumorigenesis. Surgery, 161(1), 257-263.
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