Dab solution

Manufactured by Vector Laboratories

Sourced in United States, United Kingdom

The DAB solution is a chromogenic substrate used for the detection of peroxidase enzyme activity in immunohistochemistry and other applications. It produces a brown-colored precipitate at the site of the peroxidase label, allowing for the visualization of the target antigen.

Automatically generated - may contain errors

Lab products found in correlation

Streptavidin horseradish peroxidase conjugate, by Vector Laboratories (6 mentions) Abc reagent, by Vector Laboratories (6 mentions) Alcohol solutions, by Merck Group (3 mentions) Citrate buffer, by Teknova (3 mentions) Biotinylated anti mouse igg, by Vector Laboratories (3 mentions) Avidin biotin hrp complex, by Vector Laboratories (3 mentions) Bovine serum albumin (bsa), by Roche (2 mentions) Goat anti rabbit igg conjugated to horseradish peroxidase, by Abcam (2 mentions) Rabbit anti human tenascin c monoclonal antibody, by Abcam (2 mentions) Cytoseal xyl mounting media, by Thermo Fisher Scientific (2 mentions) Axio imager a2, by Zeiss (2 mentions) Abc solution, by Vector Laboratories (2 mentions) Abc kit, by Vector Laboratories (2 mentions) Mayer s hematoxylin, by Merck Group (2 mentions) Brn801a, by Tissue Array (1 mentions) Bc081120, by Tissue Array (1 mentions) Nox2 primary antibody, by Proteintech (1 mentions) Sc 2357, by Santa Cruz Biotechnology (1 mentions) Peroxidase conjugated secondary antibody, by Vector Laboratories (1 mentions) Anti il6 antibody, by Proteintech (1 mentions)

46 protocols using dab solution

Immunohistochemistry and Prussian Blue Staining Protocol

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Immunohistochemical staining was performed using a standard protocol. Briefly, sections were allowed to dry and were permeabilized for 10 min in 1% Triton X-100 in PBS, followed by blocking in 2% BSA (Roche) and overnight staining at 4°C with the primary antibodies diluted in blocking solution. After endogenous peroxidase inhibition with 3% H₂O₂, sections were incubated with appropriate biotin-conjugate secondary antibody for 1 h at room temperature. Following signal amplification with avidin–biotin-complex–HRP (VECTASTAIN), DAB solution (VECTOR) was applied for 2–3 min. Haematoxylin (Bioptica) was further used to stain nuclei. Haematoxylin and 1% eosin were used for the H&E staining using a standard protocol.
Prussian blue staining was performed using equal parts of hydrochloric acid and potassium ferrocyanide prepared immediately prior to use, which was added to the slides for 20 min. Slides were then washed with distilled water three times and mounted in Mowiol.

Massimo M., Barelli C., Moreno C., Collesi C., Holloway R.K., Crespo B., Zentilin L., Williams A., Miron V.E., Giacca M, & Long K.R. (2023). Haemorrhage of human foetal cortex associated with SARS-CoV-2 infection. Brain, 146(3), 1175-1185.

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Immunohistochemical Analysis of Tissue Microarrays

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The paraffin-embedded tissue arrays (BRN801a and BC081120, US Biomax, Rockville, MD) were dewaxed in xylene for 10 min twice and rehydrated through a series of alcohol solutions (200 proof, Sigma-Aldrich, St. Louis, MO) (100% ethanol twice, 90% ethanol, 70% ethanol, 5 min each) to water. Antigen retrieval was done by boiling the slides for 15 min in a citrate buffer (Teknova, U.S) at pH 6.0. Endogenous peroxidase activity was blocked with 3% H₂O₂ in methanol for 10 min after returning to room temperature. The TMAs were subsequently blocked with 2% BSA and incubated with monoclonal antibody (1:75) overnight at 4°C followed by incubation with a secondary antibody conjugated to horseradish peroxidase. Immunodetection was performed using DAB solution (Vector Laboratories, Burlingame, CA). Hematoxylin counterstain was used to visualize nuclei. The protein expression level in each tissue section was assessed in non-necrotic areas of three separate microscopic fields of view under a magnification of 20× and was represented by the mean of the percentage of positive cells.

Nie S., McDermott S.P., Deol Y., Tan Z., Wicha M.S, & Lubman D.M. (2015). A quantitative proteomics analysis of MCF7 breast cancer stem and progenitor cell populations. Proteomics, 15(22), 3772-3783.

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Immunohistochemical Analysis of Liver Tissue

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Tissue was fixed by incubation in 4% formol overnight at 4 °C and embedded in paraffin wax. Hematoxylin/eosin staining was carried out on 5-µm paraffin sections. For immunohistochemistry, liver sections (5 µm) were de-paraffinized and incubated in citrate buffer at 95 °C for 20 min for antigen retrieval. Sections were treated with 3% hydrogen peroxide for 15 min at room temperature and then incubated overnight at 4 °C with the primary antibodies referenced in Table 2. After three washes in PBS1X, tissue sections were incubated with biotinylated anti-mouse/rabbit or rat IgG (1/200 dilution, Vector Laboratories, CA, USA) for 1 hr at RT and then washed three times in PBS1X, after which streptavidin–horseradish peroxidase conjugates (Vector Laboratories, CA, USA) were added and the slides incubated for 45 min. After three washes with PBS1X, DAB solution (Vector Laboratories, CA, USA) was added and the slides were counterstained with haematoxylin.

Fortier M., Cadoux M., Boussetta N., Pham S., Donné R., Couty J.P., Desdouets C, & Celton-Morizur S. (2019). Hepatospecific ablation of p38α MAPK governs liver regeneration through modulation of inflammatory response to CCl4-induced acute injury. Scientific Reports, 9, 14614.

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PCNA Immunohistochemistry for Tumor Proliferation

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Subcutaneous tumor tissues were obtained from mice under different treatments. Then the fresh tumor sections were fixed in 4% paraformaldehyde (PFA). The tumor samples were gradually dehydrated and then embedded in paraffin. The tumor sections in the paraffin were cut into 5μm in thickness and then deparaffinized and incubated in citrate buffer at 95℃ for 40 min for antigen retrieval, and then incubated overnight at 4℃with primary antibody (PCNA,1:200, CST, USA). After three washes, tissue slices were incubated with biotinylated anti-mouse IgG (1:200 dilution, CST, USA) for 1 hour at RT and then washed three times.Then streptavidinhorseradish peroxidase conjugates (Vector Laboratories, USA) were added to the slices and incubated for 45 min. After washing three times, DAB solution (Vector Laboratories, USA) was added and the slides were counterstained with hematoxylin.Proliferation index which refers to positive cells/tumor cells with brown staining were counted in 10 consecutive areas.

Qu J., Chen Q., Bing Z., Shen S., Hou Y., Lv M, & Wang T. (2023). C. tropicalis promotes CRC by down-regulating tumor cell-intrinsic PD-1 receptor via autophagy. Journal of Cancer, 14(10), 1794-1808.

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Immunohistochemical Analysis of NOX2 in Lung Tissue

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The fresh lung tissues were fixed in 4% paraformaldehyde (PFA). Then, the samples were gradually dehydrated and embedded in paraffin. Organ sections were cut into 5μm in thickness and then deparaffinized and incubated in citrate buffer at 95 °C for 40 min for antigen retrieval and then incubated overnight at 4 °C with NOX₂ primary antibody (1:100 dilution, proteintech, 19013-1-AP). After three washes, tissue slices were incubated with biotinylated anti-mouse IgG (1:200 dilution, Vector Laboratories, CA, USA) for 1 hour at RT and then washed three times, after which streptavidin-horseradish peroxidase conjugates (Vector Laboratories, CA, USA) were added and the slices incubated for 45 min. After three washes with PBS, DAB solution (Vector Laboratories, CA, USA) was added and the slides were counterstained with hematoxylin.

Sun Z., Qu J., Xia X., Pan Y., Liu X., Liang H., Dou H, & Hou Y. (2021). 17β-Estradiol promotes LC3B-associated phagocytosis in trained immunity of female mice against sepsis. International Journal of Biological Sciences, 17(2), 460-474.

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Immunohistochemistry and Immunofluorescence Staining of Brain Tissues

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Brains were fixed in 4% of PFA, flash frozen, and cut into 50 µm slices. Next, the slices were permeabilized with 3% H₂O₂, 10% methanol, and 2% of triton for 20 m. After three washes the slices were treated with the blocking solution (3% BSA, 0,1% tween 20) and incubated with the primary antibody over night at 4 °C. For the immunofluorescence the secondary antibody was added, and the slices mounted for the imaging. When immunohistochemistry was preferable, the biotinylated secondary antibody was coupled with avidin/biotinylated enzyme complex for 1 h and then revealed with the DAB solution (Vector Laboratories). For those antibodies whose function depends on antigen retrieval, the slices were treated with the antigen retrieval solution (1 mM EDTA, 0.05% Tween 20, pH 8.0) and left at 80 °C for 30 min. All the antibodies used for this study are listed in Supplementary Table 1.

Bido S., Muggeo S., Massimino L., Marzi M.J., Giannelli S.G., Melacini E., Nannoni M., Gambarè D., Bellini E., Ordazzo G., Rossi G., Maffezzini C., Iannelli A., Luoni M., Bacigaluppi M., Gregori S., Nicassio F, & Broccoli V. (2021). Microglia-specific overexpression of α-synuclein leads to severe dopaminergic neurodegeneration by phagocytic exhaustion and oxidative toxicity. Nature Communications, 12, 6237.

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Immunohistochemical Profiling of Cancer Markers

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Immunohistochemical staining for Ki67, p-Met, pSTAT3, K5/6, K18, ALDH1, p63, Vimentin w done as described previously (Burga et al., 2009 (link), Burga et al., 2011 (link)). Immunolabeling was visualized with DAB solution (Vector Laboratories), followed by counterstaining with hematoxylin.

Hu H., Luo M.L., Desmedt C., Nabavi S., Yadegarynia S., Hong A., Konstantinopoulos P.A., Gabrielson E., Hines-Boykin R., Pihan G., Yuan X., Sotirious C., Dittmer D.P., Fingeroth J.D, & Wulf G.M. (2016). Epstein–Barr Virus Infection of Mammary Epithelial Cells Promotes Malignant Transformation. EBioMedicine, 9, 148-160.

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Immunohistochemical Analysis of Tenascin-C Expression

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Immunohistochemical staining was performed using tissue microarray samples. The paraffin-embedded tissue arrays with 1.5 mm core diameter and 5 µm thickness were dewaxed in xylene for 10 min twice and rehydrated through a series of alcohol solutions (200 proof, Sigma-Aldrich, St. Louis, MO) (100% ethanol twice, 90% ethanol, and 70% ethanol, 5 min each) to water. Then, the slides were boiled for 15 min in citrate buffer (Teknova, Hollister, CA) at pH 6.0 for antigen retrieval. After returning to room temperature, endogenous peroxidase activity was blocked with 3% H₂O₂ in methanol for 10 min. The TMAs were then rinsed with water and PBS and subsequently blocked with 2% BSA and incubated with rabbit anti-human Tenascin-C monoclonal antibody (1:100 dilution, Abcam, Cambridge, MA) overnight at 4 °C followed by incubation with a goat anti-rabbit IgG conjugated to horseradish peroxidase (1:250 Abcam, Cambridge, MA). Immunodetection was performed using DAB solution (Vector Laboratories, Burlingame, CA). Hematoxylin counterstain was used to visualize nuclei. The TNC expression level in each tissue section was assessed in non-necrotic areas of three separate microscopic fields of view under a magnification of 200× and was represented by the mean of the percentage of TNC⁺ cells. The results were confirmed by a pathologist.

Nie S., Gurrea M., Zhu J., Thakolwiboon S., Heth J.A., Muraszko K.M., Fan X, & Lubman D.M. (2014). Tenascin-C: A Novel Candidate Marker for Cancer Stem Cells in Glioblastoma Identified by Tissue Microarrays. Journal of Proteome Research, 14(2), 814-822.

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Immunohistochemical Analysis of Tenascin-C Expression

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Immunohistochemical
staining was performed using tissue microarray
samples. The paraffin-embedded tissue arrays with 1.5 mm core diameter
and 5 μm thickness were dewaxed in xylene for 10 min twice and
rehydrated through a series of alcohol solutions (200 proof, Sigma-Aldrich,
St. Louis, MO) (100% ethanol twice, 90% ethanol, and 70% ethanol,
5 min each) to water. Then, the slides were boiled for 15 min in citrate
buffer (Teknova, Hollister, CA) at pH 6.0 for antigen retrieval. After
returning to room temperature, endogenous peroxidase activity was
blocked with 3% H₂O₂ in methanol for 10 min.
The TMAs were then rinsed with water and PBS and subsequently blocked
with 2% BSA and incubated with rabbit anti-human Tenascin-C monoclonal
antibody (1:100 dilution, Abcam, Cambridge, MA) overnight at 4 °C
followed by incubation with a goat anti-rabbit IgG conjugated to horseradish
peroxidase (1:250 Abcam, Cambridge, MA). Immunodetection was performed
using DAB solution (Vector Laboratories, Burlingame, CA). Hematoxylin
counterstain was used to visualize nuclei. The TNC expression level
in each tissue section was assessed in non-necrotic areas of three
separate microscopic fields of view under a magnification of 200×
and was represented by the mean of the percentage of TNC⁺ cells. The results were confirmed by a pathologist.

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Immunohistochemistry Protocol for Tissue Samples

Cited 1 time

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Immunohistochemistry was performed as previously described [72 (link), 73 (link)]. In brief, frozen sections were fixed in a methanol/acetone mixture (1 : 1) for 10 min and blocked with 10% (v/v) donkey serum in PBS at room temperature. The slides were then hybridized overnight with primary antibody or normal IgG antibody in 10% horse serum at 4°C, washed gently with PBS, and hybridized with secondary antibody conjugated to horseradish peroxidase (SC2357, Santa Cruz Biotechnology, USA) for 2 h at room temperature. The immunocomplexes were visualized using a DAB solution (Vector Laboratories, Burlingame, CA, USA). The sections were then dehydrated in a series of graded ethanol and xylene, and slides were mounted with the Cytoseal XYL mounting media (Richard-Allan Scientific, Kalamazoo, MI, USA). Finally, the sections were observed and micrographs taken with an optical microscope (Zeiss Axio Imager A2) at 10x or 40x magnification.

Yang Y., Li M., Liu Y., Wang Z., Fu X., He X., Wang Q., Li X.X., Ma H., Wang K., Zou L., Wang J.X, & Yu T. (2022). The lncRNA Punisher Regulates Apoptosis and Mitochondrial Homeostasis of Vascular Smooth Muscle Cells via Targeting miR-664a-5p and OPA1. Oxidative Medicine and Cellular Longevity, 2022, 5477024.

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