According to an existing protocol,20 immunohistochemistry was performed. The paraffin‐embedded sections were incubated with the primary antibodies (all from Abcam Inc., Cambridge, MA, USA) against interleukin (IL)‐1β (dilution ratio of 1: 200, ab9722) and IL‐18 (dilution ratio of 1: 2000, ab223293), and then incubated with the secondary antibody immunoglobulin G (IgG) (dilution ratio of 1: 2000, ab205718). Immunohistochemistry findings were analysed using the Image Pro Plus 6.0 software (Media Cybernetics, Silver Spring, USA). Five visual fields were randomly selected from each section to detect the optical density (OD) of the positive staining area. A high OD value was indicative of a high antigen expression in the positive areas.
Ab223293
Manufactured by Abcam
Sourced in United States, United Kingdom
Ab223293 is a laboratory equipment product manufactured by Abcam. It is a high-quality tool designed for researchers and scientists working in various fields. The core function of this product is to perform a specific task or procedure within the laboratory setting. Detailed information about its intended use or capabilities is not available in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Ab9722, by Abcam (2 mentions)
Ab205718, by Abcam (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Et1610 93, by Huabio (1 mentions)
Et1608 69, by Huabio (1 mentions)
Pannoramic scan, by 3DHISTECH (1 mentions)
Ab133566, by Abcam (1 mentions)
Ab125212, by Abcam (1 mentions)
Ab64693, by Abcam (1 mentions)
Gapdh antibody, by Merck Group (1 mentions)
Ab182422, by Abcam (1 mentions)
Ab7778, by Abcam (1 mentions)
Ab6308, by Abcam (1 mentions)
Ir503, by Agilent Technologies (1 mentions)
Rabbit monoclonal anti p smad2 antibody, by Cell Signaling Technology (1 mentions)
Mouse monoclonal anti tgf β1 antibody, by Santa Cruz Biotechnology (1 mentions)
Mouse monoclonal anti smad2 antibody, by Cell Signaling Technology (1 mentions)
A2547, by Merck Group (1 mentions)
Sc 130348, by Santa Cruz Biotechnology (1 mentions)
Ab79823, by Abcam (1 mentions)
4 protocols using ab223293
1
Immunohistochemical Analysis of IL-1β and IL-18
2
Immunohistochemical Analysis of Inflammasome Proteins
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The heart was fixed with 10% neutral formalin, embedded in paraffin, and cut into 4 μm slices. Immunohistochemistry was performed according to the procedure of the DAB detection kit (GK600505; GenenTech, Shanghai, China). In conclusion, sections were incubated with rabbit gasdermin D Antibody (1 : 200, 93709, Cell Signaling Technology, USA), rabbit NLRP3 antibody (1 : 200, ET1610-93, HUABIO, China), rabbit caspase-1 antibody (1 : 200, ET1608-69, HUABIO, China), rabbit anti-IL-1beta antibody (1 : 200, ab9722, Abcam, UK), and rabbit anti-IL-18 antibody (1 : 1000, ab223293, Abcam, UK) and detected by a DAB colorimetric detection kit. Then, the nuclei were stained with hematoxylin. Finally, the images were obtained using a digital slide scanner (Pannoramic SCAN, 3DHISTECH, Budapest, Hungary) and analyzed using ImageJ as described previously [29 (link)]. The protein contents were quantified by the average grey value (immunostaining intensity) and the percentage of positive area (staining area) of the positive cells [29 (link)].
3
Immunohistochemical Characterization of Inflammatory Markers
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As primary antibodies, we used a rabbit monoclonal anti-HMGB1 antibody (ab79823; Abcam, Cambridge, UK), rabbit monoclonal anti-IL-18 antibody (ab223293; Abcam), rabbit monoclonal anti-IFN-γ antibody (ab133566; Abcam), mouse monoclonal anti-α-SMA antibody (A-2547; Sigma-Aldrich; St. Louis, MO), mouse monoclonal anti- TGF-β1 antibody (sc-130348; Santa Cruz Biotechnology, Santa Cruz, CA), mouse monoclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (G8795; Sigma-Aldrich), rabbit monoclonal anti-p-Smad2 antibody (#3108; Cell Signaling Technology, Danvers, MA), mouse monoclonal anti-Smad2 antibody (#3103; Cell Signaling Technology), rabbit polyclonal anti-Col-I antibody (ab6308; Abcam), rabbit polyclonal anti-Col-III antibody (ab7778; Abcam), rabbit polyclonal anti-CD3 antibody (IR503; Dako, Santa Clara, CA), rabbit polyclonal anti-CD68 antibody (ab125212; Abcam), rabbit polyclonal anti-CD206 antibody (ab64693; Abcam) and rabbit monoclonal anti CD163 antibody (ab182422; Abcam).
4
Immunohistochemical Analysis of Inflammatory Markers in Rat Prostate
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The rat prostate slices were de-waxed in xylene prior to ethanol gradient dehydration. Sections were received 10mM citrate buffer (pH, 6.0) for antigen retrieval and immersed in 3% hydrogen peroxide to block the endogenous peroxidase before immunohistochemistry (IHC). Sections were incubated with the primary antibodies against Ki-67 (2 µg/mL, ab15580, Abcam, Cambridge, MA, USA), IL-6 (1:400, ab6672, Abcam), IL-18 (1:2,000, ab223293, Abcam), and TNF-α (1:200, ab270264, Abcam) overnight at 4°C. Sections were washed with phosphatebuffered saline (PBS) thrice before and after incubation with the secondary antibody. After that, 1–3 minutes of diaminobenzidine color development was terminated. The nucleus of cells was stained by hematoxylin for 3 minutes, and sections were dehydrated, permeabilized, and sealed. Images were acquired using a light microscope (Nikon Eclipse E100, Tokyo, Japan).
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