Diaminobenzidine dab

Manufactured by Solarbio

Sourced in China

Diaminobenzidine (DAB) is a chromogenic substrate used in immunohistochemistry and other applications to produce a brown stain. It serves as an electron donor, reacting with peroxidase-labeled antibodies to create a visible precipitate.

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Lab products found in correlation

In situ cell death detection kit, by Roche (2 mentions) Hematoxylin, by Solarbio (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Hpa005565, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Solarbio (1 mentions) Citric acid, by Solarbio (1 mentions) Ckx53, by Olympus (1 mentions) Ab75875, by Abcam (1 mentions) Goat anti rabbit igg antibody, by Affinity Biosciences (1 mentions) Normal goat serum (ngs), by Solarbio (1 mentions) Anti nf κb p65, by Wanlei (1 mentions) Dp73 optical microscope, by Olympus (1 mentions) Mounting medium, by Solarbio (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions) One step tunel apoptosis assay kit, by Beyotime (1 mentions) Triton x 100, by Beyotime (1 mentions) Anti ki67 antibody, by Solarbio (1 mentions) Goat serum, by Solarbio (1 mentions) Proteinase k, by Solarbio (1 mentions) Aluminum hydroxide gel, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

DAB (85 citations) Diaminobenzidine (DAB) developer (6 citations) Diaminobenzidine (DAB) kit (4 citations) Diaminobenzidine (DAB) solution (3 citations) 3,3′-diaminobenzidine (DAB) solution (3 citations) 3,3′-diaminobenzidine(DAB) substrate kit (2 citations) Diaminobenzidine (DAB) chromogenic kit (2 citations) HRP‐conjugated secondary antibody and diaminobenzidine (DAB) solution (2 citations) DAB (3,3′-diaminobenzidine) (2 citations)

17 protocols using diaminobenzidine dab

Immunohistochemical Analysis of Ki67

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The
immunohistochemistry of Ki67 was carried out as described in ref (38 (link)). Anti-Ki67 antibody was
used at a dilution of 1:2000, and then goat antirabbit secondary antibody
was used. Immunoreactions were detected with diaminobenzidine (DAB;
Solarbio). Images were captured and the Ki67-positive cells were counted.

Lv C., Li S., Zhao J., Yang P, & Yang C. (2022). M1 Macrophages Enhance Survival and Invasion of Oral Squamous Cell Carcinoma by Inducing GDF15-Mediated ErbB2 Phosphorylation. ACS Omega, 7(13), 11405-11414.

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Immunohistochemical Analysis of CDCA7 Expression

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Briefly, tumor tissues were fixed and cut into 4-μm-thick sections. Then, sections were deparaffinized in xylene and dehydrated in a graded ethanol series. Next, sections were immersed in sodium citrate and heated for antigen retrieval. After incubation with 3% H₂O₂ methanol solution for 15 min, sections were blocked with 10% goat serum and then incubated overnight at 4°C with anti-CDCA7 (Sigma-Aldrich, HPA005565, 1:50). The secondary antibody was used to incubate the sections for 20 min at room temperature. Diaminobenzidine (DAB; Solarbio, Beijing, China) was applied for staining development and sections were counterstained with hematoxylin for 10 min. The staining results were observed under a light microscope (Leica, Wetzlar, Germany).

Cai C., Peng X, & Zhang Y. (2021). Downregulation of cell division cycle-associated protein 7 (CDCA7) suppresses cell proliferation, arrests cell cycle of ovarian cancer, and restrains angiogenesis by modulating enhancer of zeste homolog 2 (EZH2) expression. Bioengineered, 12(1), 7007-7019.

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Immunohistochemical Staining of Colon Tissue

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After dewaxing the paraffin-embedded colon sections, citric acid (Solarbio, China) was used for antigen reparation. Then, the slices were rinsed with 3% H₂O₂ (Solarbio, China) for 25 min at room temperature to block endogenous peroxidase activity and incubated with 3% bovine serum albumin (Solarbio, China) for 30 min to block nonspecific binding. Followed by setting with the first antibodies (Proteintech, USA) in a wet box at 4°C overnight, the sections were incubated with the corresponding secondary antibody: anti-rabbit IgG antibody conjugated to horseradish peroxidase (1 : 200) (Proteintech, USA). Then, the freshly prepared diaminobenzidine (DAB) (Solarbio, China) chromogenic solution was dripped on the slices for coloration. The coloration time was controlled under a microscope (CKX53, Olympus, Japan), rinsing with tap water to stop. Counterstaining was performed using HE (Solarbio, China), sealing the sections with synthetic resin (Servicebio, China). Finally, we conducted the microscope examination and the positive results were shown in brown.

Wang L, & Wang J. (2022). Honokiol Ameliorates DSS-Induced Mouse Colitis by Inhibiting Inflammation and Oxidative Stress and Improving the Intestinal Barrier. Oxidative Medicine and Cellular Longevity, 2022, 1755608.

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Quantifying Substantia Nigra Tyrosine Hydroxylase

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Substantia nigra Sect. (5 μm) from each group were dewaxed and dehydrated. Then, the sections were treated with 3% BSA for 1 h at 25 °C and incubated with rabbit anti-mouse tyrosine hydroxylase (TH) (1:100, ab75875, Abcam, UK) overnight at 4 °C. After washing the sections with PBS, they were incubated with goat anti-rabbit IgG antibody (1:500, s0001, Affinity, AUS) at room temperature for 2 h, stained with diaminobenzidine (DAB, Solarbio, China), and counter-stained with hematoxylin. The TH-positive area of the substantia nigra was observed under a microscope, and five fields were randomly selected for image analysis. Immunohistochemical images were quantified using Image J (NIH, USA) analysis software. Mean density, which represents the expression of protein, is the cumulative integrated optical density (IOD) divided by the area of the effective target distribution.

Li M., Zhang J., Jiang L., Wang W., Feng X., Liu M, & Yang D. (2023). Neuroprotective effects of morroniside from Cornus officinalis sieb. Et zucc against Parkinson’s disease via inhibiting oxidative stress and ferroptosis. BMC Complementary Medicine and Therapies, 23, 218.

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Immunohistochemical Analysis of Rat Kidney

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After the paraffin sections with a diameter of 1 cm and a thickness of 4 μm were deparaffinized, the rat kidney tissues were incubated with 3% H₂O₂ solution for 25 min. After rinsing the tissues with PBS, the tissues were blocked with 10% normal goat serum (Solarbio Science & Technology Co., Ltd., Beijing, China) at room temperature for 1 h, and then incubated with the primary antibody (MAP4) at 4°C overnight. Subsequently, an anti-rabbit IgG antibody (diluted 1:500) was used to react with the tissues at room temperature for 2 h. The tissues were then reacted with diaminobenzidine (DAB, Solarbio Science & Technology Co., Ltd., Beijing, China) for 10 min. Finally, images were obtained through microscope.

Guan T., Zheng Y., Jin S., Wang S., Hu M., Liu X., Huang S, & Liu Y. (2022). Troxerutin alleviates kidney injury in rats via PI3K/AKT pathway by enhancing MAP4 expression. Food & Nutrition Research, 66, 10.29219/fnr.v66.8469.

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Immunohistochemical Analysis of NF-κB (P65) in Xenograft Tissue

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The expression of NF-κB (P65) in the paraffin-embedded xenograft tissue was detected by immunohistochemistry. Tissues were incubated with the anti-NF-κB (P65) (1:200; Wanleibio) primary antibody at 4 °C overnight and then with the secondary antibody at 37 °C for 60 min and an HRP-labelled streptavidin solution for 10 min. Next, tissues were stained by diaminobenzidine (DAB) (Solarbio, China) and observed under a microscope. The mean density of P65 in the tissues was calculated using Image-Pro software.

Shi Y, & Liu C. (2021). Circular RNA hsa_circ_0043278 inhibits breast cancer progression via the miR-455-3p/EI24 signalling pathway. BMC Cancer, 21, 1249.

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TUNEL Assay for Apoptosis Detection

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Excised tumor tissues were fixed in 4% paraformaldehyde, paraffin embedded, and sectioned. Some sections were stained with hematoxylin and eosin (H&E) for histological examination following the standard protocol, and some sections were subjected to TUNEL assay. TUNEL staining was performed with the In Situ Cell Death Detection Kit (Roche, Basel, Switzerland). Sections were permeabilized with 0.1% v/v Triton-X 100 for 8 min at RT. Endogenous peroxidase was blocked by incubating the sections with 2% H₂O₂ for 10 min. Sections were incubated with the enzyme solution and TUNEL reaction solution for 1 h at 37°C, followed by incubation with Converter-POD for 30 min at 37°C. Thereafter, Diaminobenzidine (DAB) (Solarbio) was added for a brief incubation of 3 to 5 min followed by counterstaining with hematoxylin for the nuclei. The sections were mounted and observed under an Olympus DP73 optical microscope.

Wang H., Sun R., Gu M., Li S., Zhang B., Chi Z, & Hao L. (2015). shRNA-Mediated Silencing of Y-Box Binding Protein-1 (YB-1) Suppresses Growth of Neuroblastoma Cell SH-SY5Y In Vitro and In Vivo. PLoS ONE, 10(5), e0127224.

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Tissue Immunohistochemistry Protocol

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For histological analysis,
the tissues were fixed in 4% formaldehyde for IHC analysis. In brief,
the slides (5 μm) were dewaxed using xylene and rehydrated using
graded alcohols. Antigen retrieval was performed with sodium citrate
(pH = 6) buffer at 92–99 °C for 15 min, and then the slides
were cooled at room temperature. The slides were washed three times
for 5 min in PBS and incubated with 3% H₂O₂ for
30 min to block endogenous peroxidases. The slides were blocked using
1% bovine serum albumin (BSA) for 30 min at room temperature. Next,
the slides were incubated with specific primary antibodies at 4 °C
overnight before being incubated using biotin-labeled secondary antibody
for 1 h at 37 °C and incubated again with diaminobenzidine (DAB)
(Solarbio Science & Technology, Beijing, China). Finally, the
slides were counterstained using hematoxylin and mounted. All experiments
were repeated independently at least three times.

Xiong J., Guo G., Guo L., Wang Z., Chen Z., Nan Y., Cao Y., Li R., Yang X., Dong J., Jin X., Yang W, & Huang Q. (2021). Amlexanox Enhances Temozolomide-Induced Antitumor Effects in Human Glioblastoma Cells by Inhibiting IKBKE and the Akt-mTOR Signaling Pathway. ACS Omega, 6(6), 4289-4299.

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TUNEL Assay for Apoptosis Detection

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Fixed cell samples were treated with 0.1% TritonX-100 (Beyotime), followed by incubating with prepared TUNEL detective solution based on the One Step TUNEL Apoptosis Assay Kit (Beyotime). After counter-staining by DAPI (Beyotime), cells were added with mounting medium (Solarbio) and finally checked and pictured under a microscope (400x) (Olympus). Tumor tissues were embedded into paraffin and cut into 5 μm slices. After de-waxing and rehydrating, TritonX-100 (0.1%, Beyotime) was used for permeabilization of slices, followed by PBS washing and TUNEL reaction through an In Situ Cell Death Detection Kit (Roche, Switzerland). Diaminobenzidine (DAB, Solarbio) and hematoxylin (Solarbio) were added to slices for coloration and counter-staining, respectively. The slices were observed under a microscope (Olympus) at a 400x magnification after mounting.

Liu H., Chi Z., Jin H, & Yang W. (2021). MicroRNA miR-188-5p as a mediator of long non-coding RNA MALAT1 regulates cell proliferation and apoptosis in multiple myeloma. Bioengineered, 12(1), 1611-1626.

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Immunohistochemical Analysis of Ki67 in Tumor Tissues

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The tumor tissues fixed in 4% paraformaldehyde (at room temperature for 24 h) were embedded in paraffin and then sectioned into 5-µm slices. After dewaxing, rehydration, and antigen retrieval, the tumor slices were incubated with 3% H₂O₂ at room temperature for 15 min. Following 15 min of blocking with goat serum (Beijing Solarbio Science & Technology Co., Ltd.) at room temperature, tumor slices were incubated with a anti-Ki67 antibody (1:100 dilution; cat. no. A2094; ABclonal Biotech Co., Ltd.) overnight at 4°C. Subsequently, the sections were incubated with an HRP-conjugated secondary antibody (cat. no. 31460; Thermo Fisher Scientific, Inc.) at 37°C for 1 h. The sections were then stained with diaminobenzidine (DAB; Beijing Solarbio Science & Technology Co., Ltd.) at room temperature for 5 min. Following DAB visualization, the sections were immersed into water and then re-stained using hematoxylin at room temperature for 3 min. The Ki67-positive cells were finally observed under a light microscope (BX53; Olympus Corporation).

Zhang C., Wang R., Li M, & Yang Q. (2023). Long non‑coding RNA BLACAT2/miR‑378a‑3p/YY1 feedback loop promotes the proliferation, migration and invasion of uterine corpus endometrial carcinoma. Oncology Reports, 49(5), 108.

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