Diaminobenzidine dab

Manufactured by Biocare Medical

Sourced in United States

Diaminobenzidine (DAB) is a chromogenic substrate used in immunohistochemistry and in situ hybridization techniques. It is commonly used to detect the presence of specific proteins or nucleic acid sequences in biological samples.

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Lab products found in correlation

Target retrieval solution, by Agilent Technologies (2 mentions) Diva decloaker 20x, by Biocare Medical (1 mentions) Decloaking chamber nxgen, by Biocare Medical (1 mentions) Peroxidazed 1, by Biocare Medical (1 mentions) Rodent block m, by Biocare Medical (1 mentions) Rodent hrp polymer, by Biocare Medical (1 mentions) Mannose receptor cd206, by Abcam (1 mentions) Arginase 1 arg 1, by Proteintech (1 mentions) Mcp 1, by Abcam (1 mentions) Background sniper, by Biocare Medical (1 mentions) Horseradish peroxidase polymer, by Biocare Medical (1 mentions) Triton x 100, by Merck Group (1 mentions) Bx53 microscope, by Olympus (1 mentions) Sc 32251, by Santa Cruz Biotechnology (1 mentions)

5 protocols using diaminobenzidine dab

Immunohistochemical Staining of Sialic Acid and PECAM-1

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Tissue was deparaffinized for 20 min at 60 °C; Diva Decloaker, 20X (Biocare medical, CA, USA) and Decloaking chamber™ NXGEN were used for recovery. The sections were incubated with primary antibody (Polyclonal Antibody to Sialic Acid, abx 100,414, abbexa Ltd. Cambridge, UK), and PECAM-1 (Polyclonal Antibody Abbiotec 250,590) diluted in blocking serum and incubated for 48 h at 4 °C, then washed 3 times with TBST, for 3 min each. They were then incubated with MACH 2 Double Stain 1 polymer (Biocare Medical MRCT523, CA, USA) for 30 min and washed with TBST. Diaminobenzidine DAB (Biocare Medical, CA, USA) was used for development. A negative control, which had no primary antibody, was performed in all groups, and salivary gland tissue was used as control tissue. IHC quantification of sialic acid was performed with ImageJ software (https://www.rsbweb.nih.gov/ij) developed by the National Institute of Health (NIH), using the IHC Profiler plug-in for the quantitative analysis of immunohistochemistry samples [16 (link)].
For evaluation of CD31/PECAM-1 immunohistochemical staining, slides were observed under a light microscope, and positivity was determined in ten microvessels. [17 ]

Hernández-Jiménez C., Martínez-Cortés J., Olmos-Zuñiga J.R., Jasso-Victoria R., López-Pérez M.T., Díaz-Martínez N.E., Alonso-Gómez M., Guzmán-Cedillo A.E., Baltazares-Lipp M., Gaxiola-Gaxiola M., Méndez-Bernal A., Polo-Jeréz A., Vázquez-Minero J.C., Hernández-Pérez O, & Fernández-Solís C.O. (2023). Changes in the levels of free sialic acid during ex vivo lung perfusion do not correlate with pulmonary function. Experimental model. BMC Pulmonary Medicine, 23, 326.

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Immunohistochemical Analysis of Lung Tissue

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Lung sections were deparaffinized and antigen retrieval was performed by heating sections to 90°C for 20 min in Target Retrieval Solution (DAKO, Carpinteria, CA) using a water bath. Endogenous peroxidases were quenched using Peroxidazed-1 (Biocare Medical, Concord, CA). To block non-specific binding, sections were next incubated with Rodent Block M (Biocare Medical) for 20 min at room temperature. Sections were then incubated with antibodies to either Mannose Receptor (CD206) (1:2500, Abcam, Cambridge, UK) overnight at 4°C, Arginase-1 (Arg-1) (1:300, Proteintech Group, Chicago, IL) for 45 min at room temperature, or with monocyte chemotactic protein-1 (MCP-1) (1:1000, Abcam) overnight at 4°C, all diluted in Renaissance Diluent (Biocare Medical). Sections were then rinsed and Rabbit on Rodent HRP polymer (Biocare Medical) was next applied for 25 min at room temperature and binding was visualized using diaminobenzidine (DAB) (Biocare Medical).

Groves A.M., Johnston C.J., Misra R.S., Williams J.P, & Finkelstein J.N. (2016). Effects of IL-4 on Pulmonary Fibrosis and the Accumulation and Phenotype of Macrophage Subpopulations Following Thoracic Irradiation. International journal of radiation biology, 92(12), 754-765.

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Immunohistochemical Staining of 5T4 Antigen

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Formalin-fixed, paraffin-embedded tissue sections were deparaffinized in xylene and rehydrated in graded ethanol. Antigen retrieval was carried out in Target Retrieval Solution (Dako). The tissue was permeabilized in 0.1% Triton-x-100 (Sigma) for 20 minutes. Following blocking with Background Sniper (Biocare Medical), tissue sections were exposed to rabbit anti-5T4 (Abcam #134162) at 4°C overnight. Tissue sections were then labeled with MACH3 probe (Biocare Medical), followed by exposure to Horseradish Peroxidase Polymer (Biocare Medical) and visualization with diaminobenzidine (DAB; Biocare Medical).

Kerk S.A., Finkel K.A., Pearson A.T., Warner K.A., Zhang Z., Nör F., Wagner V.P., Vargas P.A., Wicha M.S., Hurt E.M., Hollingsworth R.E., Tice D.A, & Nör J.E. (2016). 5T4-targeted therapy ablates cancer stem cells and prevents recurrence of head and neck squamous cell carcinoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(10), 2516-2527.

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Immunohistochemical Analysis of OSM in Breast Cancer

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Formalin-fixed, paraffin-embedded (FFPE) tissues were retrospectively collected from patients with histologically confirmed invasive ductal luminal (n = 40) or triple-negative (n = 40) breast cancers, who underwent surgery at Humanitas Clinical and Research Institute (Rozzano-Milan, Italy). FFPE sections (3 μm) were incubated with an anti-OSM antibody (Sigma-Aldrich, Milan, Italy) for 1 h at room temperature. After washing with phosphate buffered saline (PBS), MACH 1 Universal horseradish peroxidase (HRP) Polymer (Biocare Medical, Concord, CA, USA), and diaminobenzidine (DAB; Biocare Medical) were used for chromogenic immunodetection, followed by counterstaining with hematoxylin. Negative control slides without primary antibody and positive controls were used for each immunostaining run. Images were captured using an Olympus BX53 microscope (Olympus, Tokyo, Japan). Semi-quantitative analysis of the staining intensity score was performed as previously described [40 (link)]. Briefly, the staining intensity was assessed on a scale from 0 to 3 (0, negative; 1, weak; 2, moderate; 3, strong), and staining percentage was scored from 0 to 4 (0, 0%–5% staining; 4, 75%–100% staining). A composite score was then calculated by multiplying the staining intensity by the staining percentage.

Bottai G., Diao L., Baggerly K.A., Paladini L., Győrffy B., Raschioni C., Pusztai L., Calin G.A, & Santarpia L. (2017). Integrated MicroRNA–mRNA Profiling Identifies Oncostatin M as a Marker of Mesenchymal-Like ER-Negative/HER2-Negative Breast Cancer. International Journal of Molecular Sciences, 18(1), 194.

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Immunohistochemical Analysis of Smooth Muscle and Proliferation

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Paraffin sections were submitted to antigen retrieval in citrate buffer, pH 6.0 at 92°C for 40 min and treated in 3% 471 Reproduction (2020) 160 469-480 H 2 O 2 -methanol solution to block endogenous peroxidase. The elimination of unspecific binding was made by background sniper blocker (Biocare Medical, Concord, CA, USA) for 15 min. Sections were incubated overnight at 4°C with the following primary antibodies: mouse anti-human α-smooth muscle actin (sc-32,251, Santa Cruz Biotechnology) and mouse anti-human proliferating cell nuclear antigen (PCNA; mouse anti-human, sc-56, Santa Cruz Biotechnology). Both antibodies were diluted 1:100 in 1% BSA. The sections were washed in PBST and incubated with Universal Link secondary antibody for 45 min. The antigen-antibody complexes were detected by Streptavidin-HRP polymer (Star Trek Universal HRP Detection System-Biocare Kit). The reactions were revealed by diaminobenzidine -DAB (Biocare) and sections were counterstained with hematoxylin. The relative frequency of smooth muscle cells was determined using the Weibel's method described above. PCNA-positive cells were visually counted at 40× magnification, using ten microscopic fields from each histological section (4 animals, 40 microscopic fields per group).

Cassimiro I.S., Cruz A.R., Bosque B.P., de Melo Gomes L.C., Zanon R.G., da Costa Silva J.R., Fujimura P.T., Ueira-Vieira C, & Ribeiro D.L. (2020). Rat postnatal prostate development is impaired by in vitro high-glucose environment. Reproduction (Cambridge, England), 160(3).

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