Following euthanasia, the colons of mice were fixed in carnoy fixation overnight at room temperature and then embedded in paraffin. Tissues were sectioned at 5 μm thickness, and blocked with 5% BSA (Solarbio, China) for 1 h at room temperature and then incubated with primary antibodies overnight at 4°C and incubated with fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody (ZSGB Bio, China) for 1 h at room temperature, then stained by DAB Staining Kit (ZSGB Bio, China), followed by hematoxylin nuclear counterstaining. The primary antibody of rabbit anti-MUC2 (Abcam, United States) was used.
Dab staining kit
Manufactured by ZSGB-BIO
Sourced in China
The DAB staining kit is a laboratory tool used to detect specific proteins or molecules within cells or tissues through a chromogenic reaction. The kit contains the necessary reagents and solutions to perform this staining procedure, which produces a brown color at the target sites. The core function of the DAB staining kit is to enable visualization and analysis of the distribution and localization of the target analytes in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti phospho jnk tyr183 tyr185, by Cell Signaling Technology (2 mentions)
Sp9001 kit, by ZSGB-BIO (2 mentions)
Anti jnk, by Cell Signaling Technology (2 mentions)
Anti gm130, by BD (2 mentions)
Nissl staining solution, by Beyotime (2 mentions)
Bovine serum albumin (bsa), by Solarbio (1 mentions)
Bovine serum albumin (bsa), by Jackson ImmunoResearch (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rabbit anti gfap antibody, by Merck Group (1 mentions)
Sc 137006, by Santa Cruz Biotechnology (1 mentions)
Sp kit, by ZSGB-BIO (1 mentions)
Inverted microscope, by Olympus (1 mentions)
Osteocalcin, by Abcam (1 mentions)
Balb c nude nu nu mice, by Jackson ImmunoResearch (1 mentions)
Proliferating cell nuclear antigen (pcna), by Proteintech (1 mentions)
Mecamylamine hydrochloride, by Merck Group (1 mentions)
Atropine sulfate salt monohydrate, by Merck Group (1 mentions)
Camkii, by GeneTex (1 mentions)
Anti ht7, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
20 protocols using dab staining kit
1
Immunohistochemical Analysis of Mucosal MUC2
2
Immunohistochemical Analysis of FFPE Samples
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FFPE (formalin-fixed, paraffin-embedded) sections acquired from patients or mice tissues were deparaffinated, dehydrated and subjected to antigen retrieval by placing them in citrate buffer in a 98°C water bath for 10 min to shelter endogenous peroxidase activity. The samples were also subjected to H2O2 treatment and blocked with 5 % BSA solution for 30 min. Thereafter, the samples were incubated with specific primary antibodies overnight at 4°C. On the following day, sections were incubated with corresponding secondary antibodies and then analyzed using a DAB staining kit (ZSGB-BIO).
3
Isolation and Characterization of Spinal Cord Astrocytes
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Neonatal (Sprague Dawley, female, specific-pathogen-free grade) rats (Kunming Institute of Zoology, Cas, China) were decapitated, and spinal cord samples were collected. The pia mater was pierced by blood vessels to the brain and spinal cord, and its capillaries nourished the brain. The spinal cord cells were harvested in Dulbecco’s Modified Eagle’s medium (GibcoTM, cat: 11965092, New York, NY, USA), containing 10% fetal bovine serum (Gibco™, cat: 10099141, New York, NY, USA) and 2 mmol/L of Gln (Sigma-Aldrich, cat: 1.00289, St. Louis, MO, USA). Then, the astrocytes, fibroblasts, neuron cells, oligodendroglia cells, and microglial cells were separated by centrifugation. The astrocytes were collected, blocked with bovine serum albumin (Jackson Immuno Research Laboratories Inc., cat: 001-000-161, West Grove, PA, USA), and incubated with rabbit anti-GFAP antibody (Sigma-Aldrich, cat: SAB5600060, St. Louis, MO, USA). The astrocytes store in a refrigerator at −80 ℃. Next, the astrocytes were stained with a 3,3'-diaminobenzidine (DAB) Staining Kit (ZSGB-BIO Co., Ltd., cat: ZLI-9018, Beijing; Standard Compound Microscope System, Olympus, Tokyo, Japan).
4
Immunofluorescence and Immunohistochemistry Protocols
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The slides were dewaxed in xylene, rehydrated, and then subjected to antigen retrieval in citrate buffer at 99°C for 5 min×3. For IF, the primary antibody anti-GM130 (Golgi matrix protein of 130kDa) (1:10, BD Pharmingen, USA) was incubated overnight at 4°C. Nuclear was stained using DAPI. For IHC, the primary antibodies, anti-phospho-JNK (Tyr183/Tyr185) or anti- JNK (1:50, Cell Signaling Technology Inc., MA, USA) were incubated overnight at 4°C; next procedures were performed with SP9001 kit and DAB Staining Kit (ZSGB-Bio, Beijing, China).
5
Immunohistochemical Staining of NPC Tissues
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The paraffin-embedded NPC tissues were sectioned into 5 µm-thick slides. The slides were deparaffinized, performed for antigen retrieval, and blocked with a proper blocking solution. The slides were then incubated with the primary antibody against TBL1X (1:50, #sc137006, Santacruz) or Flot2 (1:50, #28208-1-AP, Proteintech). After that, the slides were incubated with relative secondary antibodies and stained with a DAB staining kit (#ZLI9017, ZSGB BIO, Beijing, China).
6
CD8+ T Cell Immunohistochemistry
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The tissue slides of HCC sample through deparaffinization and dehydration were incubated with anti-CD8 primary antibody overnight at 4 °C after epitope retrieval, H2O2 treatment and non-specific antigens blocking. Slides were next incubated with secondary antibody, followed by signal detection with DAB staining kit (Zsbio, China)
7
Immunohistochemical Analysis of Retinal Tissues
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Dewaxed and antigen-repaired retinal paraffin sections were processed by Endogenous Peroxidase Blocking Solution (BOSTER, China) in the dark at RT for 10 minutes and blocked by goat serum at RT for 2 hours. The sections were incubated with primary antibodies (see Supplementary Table S1 ) at 37°C for 2 hours and with species-specific secondary antibodies (see Supplementary Table S1 ) at RT for 25 minutes, followed by staining with DAB Staining Kit (ZSGB-BIO, China). After redyeing with hematoxylin and dehydration with graded alcohol, we applied coverslips to the sections with neutral resin and observed them by a light microscope.
8
Immunohistochemical Analysis of PDX Tissues
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Immunohistochemistry was performed on PDX tissues by subjecting 4-µm paraffin sections to a three-step process and a DAB staining kit (ZSGB-BIO). In brief, formalin-fixed, paraffin-embedded tissue sections were dried at 80 °C for 15 min and dewaxed in xylene, rinsed in graded ethanol, and rehydrated in double-distilled water. For antigen retrieval, slides were pretreated by steaming them in sodium citrate buffer (10 mM sodium citrate, pH 6.0) for 15 min at 100 °C. After the sections were washed with phosphate-buffered saline for 3 min, they were immunostained with primary antibodies against Ki-67, TGF-β and CD34 from ZSGB-BIO and incubated at 4 °C overnight. After 3 washes in PBS buffer, the tissues were covered by HRP-conjugated anti-mouse/rabbit antibodies for 30 min at room temperature (RT). Staining reactions were performed by covering the tissue samples with the prepared DAB chromogen solution and incubating them for approximately 10 min to allow proper brown color development 24 (link).
9
Immunohistochemical Analysis of Tumor Sections
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Tumors were fixed by 4% paraformaldehyde, paraffin embed and sliced into sections, sections were hydrated by xylene and gradient alcohol (Sinoreagent, China). Antigen of sections were repaired by citrate solution and blocked by goat serum, then sections incubated with primary and secondary antibodies according to the manufacturer’s protocol of SP Kit (ZSGB-BIO, China) and stained by DAB Staining Kit (ZSGB-BIO, China) and hematoxylin solutions (Sinoreagent, China). The pictures of sections were photographed by inverted microscope (Olympus, Japan). Relative staining score was calculated using an IHC score analysis method according to the proportion of positively stained cells and the intensity of staining. The proportion of positive cells was scored as follows: 0 (0–5%), 1 (6–25%), 2 (26–50%), 3 (51–75%), 4 (> 75%) and the intensity was scored as follows: 0 (negative), 1 (weak), 2 (moderate), 3 (strong).
10
Immunohistochemical Staining of Intracranial Tumors
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IHC staining was performed on intracranial tumors by subjecting 5-μm paraffin sections to a three-step process using a DAB staining kit (Zsgb-Bio, Beijing, China), as previously reported33 (link).
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