The fixed paraffin-embedded sections of human myometrium were rehydrated in a graded series of decreasing alcohol concentrations. Sections were incubated in 3% hydrogen peroxide for 30 min to block endogenous peroxidase and 2% normal goat serum for 1 hr to reduce nonspecific binding. Then samples were incubated overnight at 4℃ with goat anti-human IL-33 monoclonal antibody (MAB36253, R&D) at a concentration of 0.1 µg/ml and then one to three drops of biotinylated secondary antibodies for 2 hr at room temperature. One to five drops of DAB chromogen solution were added so as to cover the entire tissue section and incubated for 10 min. The sections were rinsed in deionized H2O and then the slides were drained. The stained tissues were covered with a coverslip of an appropriate size and these were then visualized under a microscope using a bright-field illumination.
Mab36253
Manufactured by R&D Systems
MAB36253 is a mouse monoclonal antibody that recognizes human MCPIP1 (Monocyte Chemotactic Protein-Induced Protein 1). It is suitable for use in various immunological techniques, such as Western blot and ELISA.
Automatically generated - may contain errors
4 protocols using mab36253
1
Immunohistochemical Analysis of IL-33 in Human Myometrium
2
Immunofluorescence Staining of IL-33
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Tissue sections and cells were fixed with 4% paraformaldehyde in PBS for 30 min at room temperature, washed three times with PBS and repaired with 75% glycine for 10 min. Then they were permeabilized with 0.5% Triton X‐100 in PBS for 30 min, and then washed three times with PBS and blocked with 3% bovine serum albumin (BSA, sh30087.02, HyClone) in PBS for 1 hr. These operations were conducted at room temperature. Primary antibody incubation was performed overnight at 4°C with goat monoclonal anti-IL-33 antibody (1:200, MAB36253, R&D) diluted in 0.2% BSA in PBS. For detection, the tissues were incubated for 2 hr at room temperature with Alexa Flour 594 secondary antibody (1:500, A32758, Life Technologies) which was diluted in 0.2% BSA in PBS. The samples were washed as described above, mounted using Antifade Mounting Medium with 4',6-diamidino-2-phenylindole (DAPI) (P0131, Beyotime Biotechnology) and analyzed using a confocal fluorescence microscope (LSM 800, Zeiss).
3
Immunohistochemical Detection of IL-33 in Human Myometrium
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The fixed paraffin-embedded sections of human myometrium were rehydrated in a graded series of decreasing alcohol concentrations. Sections were incubated in 3% hydrogen peroxide for 30 minutes to block endogenous peroxidase and 2% normal goat serum for 1 hour to reduce nonspecific binding. Then samples were incubated overnight at 4 with goat anti-human IL-33 monoclonal antibody (MAB36253, R&D) at a concentration of 0.1 µg/mL and then 1-3 drops of biotinylated secondary antibodies for 2 hours at room temperature. 1-5 drops of DAB chromogen solution were added to cover the entire tissue section and incubated for 10 minutes. The sections were rinsed in deionized H2O and the slides were drained and the stained tissue was covered with a coverslip of an appropriate size. These were then visualized under a microscope using a bright-field illumination.
4
Immunofluorescence Staining of IL-33
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue sections and cells were fixed with 4% paraformaldehyde in PBS for 30 minutes at room temperature, washed three times with PBS and repaired with 75% glycine for 10 minutes. Then they permeabilized with 0.5% Triton X 100 in PBS for 30 minutes, and then washed three times with PBS and blocked with 3% bovine serum albumin (BSA, sh30087.02, HyClone) in PBS for 1 hour. These operations were conducted at room temperature. Primary antibody incubation was performed overnight at 4°C with goat monoclonal anti-IL-33 (1:200, MAB36253, R&D) diluted in 0.2% BSA in PBS. For detection, the tissues were incubated for 2 hours at room temperature with Alexa Flour 594 secondary antibody (1:500, A32758, Life Technologies) which was diluted in 0.2% BSA in PBS. The samples were washed as described above, mounted using Antifade Mounting Medium with DAPI (P0131, Beyotime Biotechnology) and analyzed using a confocal fluorescence microscope (LSM 800, Zeiss).
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