α-ZAL was a gift from Professor Shunling Dai at Perking Union Medical College. 17β-E2 and 5′-bromo-2′-deoxyuridine (BrdU) were purchased from Sigma-Aldrich (Saint Louis, MO, USA). The protease inhibitor mixture and BCA Protein Assay Kit were purchased from Pierce Biotechnology (Rockford, IL, USA). BrdU antibody was purchased from Millipore (Billerica, MA, USA). BDNF, TrkB, and p75NTR antibody were purchased from Abcam (Abcam, England); β-actin antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The avidin-biotin-peroxidase and DAB kits for immunohistochemical detection were obtained from Zhongshan Goldenbridge Biotechnology (Beijing, China). All other chemicals used were of the highest grade commercially available.
Dab kit
Manufactured by Zhongshan Golden Bridge Biotechnology
Sourced in China
The DAB kit is a laboratory reagent used for the detection and visualization of target proteins or antigens in various biological samples, such as tissue sections or cell cultures. It provides a colorimetric detection method that produces a brown reaction product, allowing for the identification and localization of the target analyte.
Automatically generated - may contain errors
Lab products found in correlation
Pv 9001, by Zhongshan Golden Bridge Biotechnology (2 mentions)
Rabbit anti human intelectin 94 145 antibody, by Phoenix Pharmaceuticals (2 mentions)
Isotype matched control rabbit igg, by Abmart (2 mentions)
Paraformaldehyde pbs, by Merck Group (2 mentions)
β actin antibody, by Santa Cruz Biotechnology (1 mentions)
Brain derived neurotrophic factor (bdnf), by Abcam (1 mentions)
Brdu antibody, by Merck Group (1 mentions)
P75ntr antibody, by Abcam (1 mentions)
5 bromo 2 deoxyuridine brdu, by Merck Group (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor mixture, by Thermo Fisher Scientific (1 mentions)
Enhanced chemiluminescence kit, by Beyotime (1 mentions)
Epigallocatechin gallate (egcg), by Yuanye Bio-Technology (1 mentions)
Nissl staining kit, by Beyotime (1 mentions)
Camkii, by Proteintech (1 mentions)
Dcfh da, by Beyotime (1 mentions)
Fluo 3 am, by Beyotime (1 mentions)
Beclin 1, by Proteintech (1 mentions)
Malondialdehyde mda kit, by Nanjing Jiancheng (1 mentions)
Ab128870, by Abcam (1 mentions)
38 protocols using dab kit
1
Reagent Preparation for Neuroscience Research
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α-ZAL was a gift from Professor Shunling Dai at Perking Union Medical College. 17β-E2 and 5′-bromo-2′-deoxyuridine (BrdU) were purchased from Sigma-Aldrich (Saint Louis, MO, USA). The protease inhibitor mixture and BCA Protein Assay Kit were purchased from Pierce Biotechnology (Rockford, IL, USA). BrdU antibody was purchased from Millipore (Billerica, MA, USA). BDNF, TrkB, and p75NTR antibody were purchased from Abcam (Abcam, England); β-actin antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The avidin-biotin-peroxidase and DAB kits for immunohistochemical detection were obtained from Zhongshan Goldenbridge Biotechnology (Beijing, China). All other chemicals used were of the highest grade commercially available.
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α-ZAL was a gift from Professor Shunling Dai at Perking Union Medical College. 17β-E2 and 5′-bromo-2′-deoxyuridine (BrdU) were purchased from Sigma-Aldrich (Saint Louis, MO, USA). The protease inhibitor mixture and BCA Protein Assay Kit were purchased from Pierce Biotechnology (Rockford, IL, USA). BrdU antibody was purchased from Millipore (Billerica, MA, USA). BDNF, TrkB, and p75NTR antibody were purchased from Abcam (Abcam, England); β-actin antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The avidin-biotin-peroxidase and DAB kits for immunohistochemical detection were obtained from Zhongshan Goldenbridge Biotechnology (Beijing, China). All other chemicals used were of the highest grade commercially available.
2
PLGA-based Neuroprotective Formulation
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mPEG-poly lactic-co-glycolic acid (PLGA) (75:25, 45,000) was purchased from Daigang Biomaterial Co., Ltd (Jinan, Shandong, China). EGCG (purity > 98%; Yuanye Biotechnology Co., Ltd, Shanghai, China) was dissolved in water (pH 3.0) and stored at −20°C. Nimodipine was obtained from Ruiyang Pharmaceutical Co., Ltd (Yi Yuan, Shandong, China). Lactate dehydrogenase (LDH), glutathione (GSH), superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) kits were obtained from the Nanjing Jiancheng Bioengineering Institute (Nanjing, Jiangsu, China). JC-1, Fluo-3 AM, DCFH-DA, Nissl staining kits, and enhanced chemiluminescence kits were purchased from Beyotime (Shanghai, China). CaMKII, Atg5, Beclin-1, and Mn-SOD antibodies were obtained from Proteintech Group, Inc (Rosemont, IL, USA). SP-9002 SPlink detection and DAB kits were purchased from Zhongshan Golden Bridge Biotechnology (Beijing, China).
3
Immunohistochemical Analysis of C19orf10 in KIRC
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Human KIRC tissue microarray including 33 cases of paraffin-embedded tissues was purchased from Shanghai Outdo Biotech Co., Ltd. The expression of C19orf10 was assessed using immunohistochemistry (IHC). The sections were dewaxed in xylene and graded alcohols, and antigen was repaired with sodium citrate solution in a microwave. Then the tissues were treated by 3% hydrogen dioxide and blocked with 5% goat serum. The sections were incubated with antibody against C19orf10 (Proteintech Group Inc., Wuhan, China), followed by incubation with secondary antibody labeled with peroxidase and visualized with a DAB kit (Zhong Shan Golden Bridge Biotechnology, Beijing, China). The nucleus was stained with hematoxylin. The IHC score was obtained by multiplying the intensity score (0, negative; 1, weak; 2, moderate; 3, strong) by the score for the percentage of positively stained cells (1, ≤25%; 2, 26%-50%; 3, 51%-75%; 4, >76% of positively stained cells).
4
Metabolic Enzyme Expression in Thyroid Tumors
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The expression of metabolic enzymes in thyroid tumor tissues was assessed via IHC staining using specific antibodies. Successive frozen tissue sections adjacent to the section analyzed using AFAIDESI-MSI were warmed to room temperature for 20 min. Sections were fixed in paraformaldehyde for 10 min. After washing in phosphate-buffered saline, the sections were immersed in 0.25% Triton X-100 for 15 min to make the tissue permeable, and then blocked with 1% bovine serum albumin for 30 min at room temperature. Furthermore, the sections were incubated with antibodies against iPLAs (Proteintech; Chicago, IL, USA; 22030-1-AP; 1:200) and FASN (Abcam; Toronto, ON, Canada, C; ab128870; 1:300) at 4 °C overnight, followed by rewarming at room temperature for 20 min. A PV-9000 two-step IHC kit was used according to the manufacturer’s instructions, and a DAB kit was used to detect antigen–antibody binding (Zhongshan Goldenbridge Biotechnology Ltd. Co., Beijing, China). The slides were counterstained with hematoxylin, dehydrated, mounted, and covered. FASN and iPLAs staining revealed cytoplasmic protein expression. The intensities of FASN and iPLAs were graded semi-quantitatively based on a scale comprising 0 (no staining), low expression (weak-to-moderate staining), and high expression (strong staining).
5
Immunohistochemical Analysis of Liver Cancer
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Paraffin-embedded liver cancer samples from patients and cancer tissue from nude mice were immunostained with antibodies after deparaffinization and hydration. The signal of IHC was amplified via a kit (#PV-9001, Zhongshan Golden Bridge Biotechnology, Beijing, China). Endogenous peroxidase was blocked with 3% hydrogen peroxide. Tissue sections were then incubated with β-catenin, YB1, AKT (pan), and ERK1/2 antibodies overnight at 4 °C. The next day, tissue sections were incubated with 100 μL or an appropriate amount of reaction enhancement solution at room temperature for 20 min. Tissue sections then received 100 μL or an appropriate amount of enhanced enzyme-labeled goat anti-rabbit IgG polymer and incubated at room temperature for 20 min. Antibody binding was assessed with a DAB kit (#ZLI-9019, Zhongshan Golden Bridge Biotechnology, Beijing, China). Then, tissue sections were counter-stained with hematoxylin. The pictures of immunostaining were captured by a microscope.
6
Paraffin Embedding of Matrigel-Organoids
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After fixed with 4% paraformaldehyde, Matrigel-organoids was aspirated from the 24-well plates into a 1.5 mL EP tube and centrifuged at 1000 rpm for 5 min, then embedded in 2% agarose. The solidified organoids-agarose was fixed in 4% paraformaldehyde overnight, and followed by gradient dehydration the next day, treated with dimethylbenzene for 40 min before embedded in paraffin. 4 μm paraffin sections were cleared with dimethylbenzene and rehydrated with gradient ethanol. For Immunohistochemistry (IHC) assay, the paraffin section antigen retrieval was performed by microwave heating and stained with the following antibodies: Rabbit anti-ER antibody (Santa, Dallas, Texas, USA, sc-8002, 1:300), anti-PR (CST, Danvers, MA, USA, 8757s, 1:600), anti-HER2 (CST, 2165s, 1:600), and anti-Ki67 (CST, 9449s, 1:800). DAB kit was purchased from Zhongshan Goldenbridge Biotechnology Company (Beijing, China). Nuclei were counterstained with haematoxylin. Images were acquired with Leica Eclipse E600 microscope.
7
Immunohistochemical Analysis of TGF-β1, Col1a1, and pSmad3
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Immunohistochemistry (IHC) was performed consistent with the manufacturer’s protocol of the universal two-step detection kit (Zhongshan Golden Bridge, Beijing, China). Slices were mounted with antibodies against TGF-β1, Col1a1, ki67 and pSmad3, and the blots were conducted with the DAB kit (Zhongshan Golden Bridge, Beijing, China). Images were taken from ×20 and ×40 objective phase contrast microscope. All control staining using PBS was provided in Supplementary Figure S1 .
8
Immunohistochemical Analysis of Kidney Proteins
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Paraffin-embedded kidney specimens were sectioned (3 μm thickness) and incubated with 3% H2O2 for 10 min. After tissue sections were washed with PBS, they were boiled in 10 mM citrate buffer (pH 6.0) for 4 min for antigen retrieval and then blocked with 10% goat serum in PBS (Zhongshan Golden Bridge Biotechnology, Beijing, China [PV-9001]) at room temperature for 1 hour. The sections were incubated with anti-KL (1:100, Abcam, Cambridge, UK [ab203576]) and anti-CASP3 (1:100, Cell Signaling Technology, Inc., Danvers, MA, USA, [9664]) at 4°C overnight. After washing, the tissue sections were incubated with HRP-conjugated secondary antibodies (Zhongshan Golden Bridge Biotechnology [PV-9001]). Color was developed using a DAB kit (Zhongshan Golden Bridge Biotechnology [ZLT-9018]). The tissue sections were counterstained with hematoxylin, dehydrated, and mounted on glass slides. Tissue sections that were incubated with secondary antibody alone were used as a negative control for IHC staining. Images at 200× magnification were acquired using a biological imaging microscope (BX53; Olympus Corporation). The intensity of positive staining, which appeared brown, was determined using Image-Pro plus 6.0 image analysis software. The mean optical density (MOD) was calculated to represent the intensity. MOD values increased as protein expression increased.
9
Immunohistochemical Analysis of Intestinal Tight Junctions and T-Cell Markers
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Paraffin-embedded intestinal tissue sections were used for immunohistochemistry. Tight junction proteins ZO-1 and Occludin expression was detected using rabbit anti-ZO-1 (1:1000, Abcam, Cambridge, MA, USA) and rabbit anti-Occludin antibodies (1:50, Abcam, Cambridge, MA, USA), respectively. Thereafter, the sections were immunostained with goat anti-rabbit IgG conjugated to horseradish peroxidase (HRP) using a DAB kit (Zhongshan Goldenbridge, Beijing, China).
T-cell markers CD3, CD4, CD8a, and CD25 were detected using mouse anti-CD3, rat anti-CD4, mouse anti-CD8a and mouse anti-CD25 (1:250, Santa Cruz Biotechnologies, Dallas, TX, USA), respectively. For the secondary antibody, we utilized goat anti-rabbit IgG conjugated to horseradish peroxidase (HRP) using a DAB kit (Vector Laboratories, Inc., Burlingame, CA, USA).
T-cell markers CD3, CD4, CD8a, and CD25 were detected using mouse anti-CD3, rat anti-CD4, mouse anti-CD8a and mouse anti-CD25 (1:250, Santa Cruz Biotechnologies, Dallas, TX, USA), respectively. For the secondary antibody, we utilized goat anti-rabbit IgG conjugated to horseradish peroxidase (HRP) using a DAB kit (Vector Laboratories, Inc., Burlingame, CA, USA).
10
Immunohistochemical Analysis of Key Proteins
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Immunoperoxidase staining for p-SAPK/JNK, DNMT1, p65, EZH2 proteins were performed on all tissues from representative subjects (control, PPI-treated groups). Briefly, the antigen was retrieved by citric acid buffer (pH 6.0) and then immersed in 3 % H2O2 to inhibit endogenous peroxidase activity, followed by incubation in 5 % bovine serum albumin to block nonspecific binding. The sections were then incubated with primary antibodies against p-SAPK/JNK (1:100), p65 (1:100), EZH2 (1:50), DNMT1 (1:100) at 4 °C overnight and then incubated with biotinylated secondary antibody (Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd. Beijing, China) for 10 min. Detection was made using the 3,3 -diaminobenzidine according to the instructions (DAB kit, Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd. China). Pictures were taken under 200× magnification. The immunostaining was evaluated by Image-Pro Plus6.0 image analysis software (Media Cybernetics, lnc. Sliver Spring, MD, USA) in at least five random high-power fields.
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