IF analysis was performed on paraffin-embedded sections. Sections were deparaffinized in xylene before being heated in citrate-EDTA antigen retrieval solution (Beyotime, China) for antigen retrieval and blocked with 1% BSA. The primary antibodies used for IF analysis were caspase-11 (Santa Cruz, USA) and GSDMD (Abcam, UK). Primary antibodies were incubated at 4 °C overnight. After being washed, the sections were incubated with Alexa Fluor 488 dye-labeled anti-rabbit IgG or Alexa Fluor 594 dye-labeled anti-mouse IgG (Beyotime, China). Finally, the sections were sealed with Antifade Mounting Medium with DAPI (Beyotime, China). Images were visualized using a confocal microscope (Olympus, Japan).
Caspase 11
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Caspase-11 is a laboratory enzyme used in research. It is a member of the caspase family of proteases, which are key regulators of apoptosis, a form of programmed cell death. Caspase-11 plays a role in the inflammatory response and can induce pyroptosis, a type of cell death. This product is intended for research use only and its specific applications should be determined by the user.
Automatically generated - may contain errors
Lab products found in correlation
Caspase 1, by Santa Cruz Biotechnology (3 mentions)
Anti gapdh, by Santa Cruz Biotechnology (2 mentions)
Psd95, by Abcam (2 mentions)
Nf κb p65, by Abcam (2 mentions)
Synapsin 1, by Merck Group (2 mentions)
Gapdh, by Abcam (2 mentions)
Il 1β, by Santa Cruz Biotechnology (2 mentions)
Pierce bca protein assay kit, by Beyotime (2 mentions)
Gsdmd, by Santa Cruz Biotechnology (2 mentions)
Il 18, by Abcam (2 mentions)
Lamin b, by Proteintech (2 mentions)
Nlrp3, by Abcam (2 mentions)
Citrate edta antigen retrieval solution, by Beyotime (1 mentions)
Gsdmd, by Abcam (1 mentions)
Antifade mounting medium with dapi, by Beyotime (1 mentions)
Catalase polyethylene glycol, by Merck Group (1 mentions)
Superoxide dismutase sod assay kit, by Nanjing Jiancheng (1 mentions)
Malondialdehyde mda assay kit, by Nanjing Jiancheng (1 mentions)
N acetyl cysteine (nac), by Merck Group (1 mentions)
Gsdmd n, by Santa Cruz Biotechnology (1 mentions)
5 protocols using caspase 11
1
Immunofluorescent Analysis of Caspase-11 and GSDMD
2
Antioxidant and Anti-inflammatory Assays
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EPZ015666 (GSK 3235025) (EPZ) was purchased from Selleck Chemicals (Houston, TX, USA). NAC and polyethylene glycol-catalase were supplied by Sigma–Aldrich (St. Louis, MO, USA). Antibodies used in western blotting (WB) and immunohistochemistry were as follows. The rabbit anti-PRMT5 and anti-GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against NLRP3, ASC, and caspase-1 and caspase-11 and GSDMD-N were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), while antibodies against Nrf2, HO-1, IL-1β, and Ki-67 were purchased from Abcam (Cambridge, UK). Dichlorofluorescein diacetate (DCFH-DA) solution was supplied by Beyotime Biotechnology (Jiangsu, China). Creatinine and urea commercial kits, the superoxide dismutase (SOD) assay kit, and the malondialdehyde (MDA) assay kit were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).
3
NAC and LY294002 Signaling Pathways
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N-acetyl-cysteine (NAC) was purchased from Sigma–Aldrich (St. Louis, MO, USA). LY294002 was supplied by Cell Signaling Technology (Danvers, MA, USA). The rabbit anti-TRIM8 and anti-GAPDH were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). These antibodies including Cleaved caspase-3, BAX, AKT, p-AKT, PI3K and p-PI3K were purchased from Cell Signaling Technology (Danvers, MA, USA), while antibodies against Bcl-2 and IL-1β were obtained from Abcam (Cambridge, UK). Primary antibodies against NLRP3, Caspase-11 and Caspase-1 and ASC and GSDMD-N were supplied by Santa Cruz Biotechnology (Santa Cruz, CA, USA).
4
Quantitative Protein Analysis via Western Blotting
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Protein quantification was performed with a Pierce BCA Protein Assay Kit (Beyotime Institute of Biotechnology), and 30–50 μg of total protein was resolved by polyacrylamide gel electrophoresis (SDS-PAGE, 8%). Protein levels were determined via incubation with antibodies against NF-κB-p65 (1:500; Abcam, UK), IκBα (1:500; Abcam, UK), NLRP3 (1:500; Abcam, UK), caspase-1 (1:500; Santa Cruz Biotechnology, USA), caspase-11 (1:500; Santa Cruz Biotechnology, USA), IL-1β (1:500; Santa Cruz Biotechnology, USA), IL-18 (1:500; Abcam, UK), Synapsin-1 (1:500; Millipore, USA), PSD-95 (1:500; Abcam, UK), GSDMD (1:500; Santa Cruz Biotechnology, USA), Lamin B (1:1000; Proteintech, USA), and GAPDH (1:500; Abcam, UK). The blots were imaged with ECL Plus western blotting detection reagents. ImageJ software was used to determine the average absorbance value of the corresponding bands.
5
Quantifying Protein Levels with Western Blot
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Protein quanti cation was performed using the Pierce BCA Protein Assay Kit (Beyotime Institute of Biotechnology), and 30-50 μg of total protein was dissolved by polyacrylamide gel electrophoresis (SDS-PAGE, 8%). Protein levels were determined via incubation with antibodies against NF-κB-p65 (1:500; Abcam, UK), IκBα (1:500; Abcam, UK), NLRP3 (1:500; Abcam, UK), caspase-1 (1:500; Santa Cruz Biotechnology, USA), caspase-11 (1:500; Santa Cruz Biotechnology, USA), IL-1β (1:500; Santa Cruz Biotechnology, USA), IL-18 (1:500; Abcam, UK), synapsin 1 (1:500; Millipore, USA), PSD-95 (1:500; Abcam, UK) GSDMD (1:500; Santa Cruz Biotechnology, USA), Lamin B (1:1000; Proteintech, USA), and GAPDH (1:500; Abcam, UK). The blots were imaged using ECL Plus western blotting detection reagents. Image J software was used to determine the average absorbance value of the corresponding bands.
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