Anti-GSDMD and anti-IL-18 were purchased from Abcam (Cambridge, UK). Anti-GAPDH, anti-cleaved Caspase 3, and anti-Caspase 3 were purchased from Cell Signaling Technology (Danvers, MA, US). Anti-Caspase 11 was purchased from Novus (Littleton, CO, USA). Anti-Caspase 1(p20) was purchased from AdipoGen (San Diego, CA, USA). Anti-HA was from Biolegend (San Diego, CA, USA). HT-29 cells were maintained in DMEM containing 10% (vol/vol) FBS, penicillin (100 U/ml) and streptomycin (100 μg/ml).
Anti caspase 11
Manufactured by Novus Biologicals
Sourced in United States
Anti-caspase-11 is a laboratory reagent used in research applications. It is a protein that specifically binds and inhibits the activity of caspase-11, a member of the caspase family of enzymes involved in programmed cell death processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti gsdmd, by Abcam (2 mentions)
Anti caspase 3, by Cell Signaling Technology (2 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Anti il 18, by Abcam (1 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
Anti ha, by BioLegend (1 mentions)
Anti caspase 1 p20, by Adipogen (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Bca protein assay, by Beyotime (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Cd11b fitc, by Thermo Fisher Scientific (1 mentions)
Myeloperoxidase mpo assay kit, by Nanjing Jiancheng (1 mentions)
Percoll, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Compound c, by Merck Group (1 mentions)
Lysis buffer, by Beyotime (1 mentions)
Anti mpo, by Abcam (1 mentions)
Anti ly6g, by Abcam (1 mentions)
Anti nrf2, by Santa Cruz Biotechnology (1 mentions)
Cxcl2, by R&D Systems (1 mentions)
5 protocols using anti caspase 11
1
Immunoblotting for Cell Death Pathway
2
Investigating Nrf2-mediated Anti-inflammatory Mechanisms
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LPS, SB225002, Percoll, and Compound C (CC) were from Sigma-Aldrich (St. Louis, MO). TMZ was from Servier (Tianjin, China). RPMI1640 medium was from Thermo Fisher Scientific (Walham, MA). Nrf2 siRNA was from RiboBio (Guangzhou, China). CXCL2 was from R&D Systems (Minneapolis, MN). DAPI and IL-1β ELISA kit were from Boster (Wuhan, China). Myeloperoxidase (MPO) assay kit and lacate dehydrogenase (LDH) assay kit were from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Lipofectamine 2000 was from Invitrogen (Waltham, MA). Protein A/G agroase, anti-Nrf2, and anti-CXCR2 were from Santa Cruz Biotechnology (Dallas, TX). Anti-Ly6G, anti-MPO, anti-GRK2, and isotype antibody were from Abcam (Cambridge, MA). Anti-AMPK and anti-phospho-AMPK were from Cell Signaling Technology (Danvers, MA). Anti-caspase-11 was from Novus Biologicals (Littleton, CO). FITC-CD11b and Percp/Cy5.5-Ly6G were from eBioscience (San Diego, CA). PE-CXCR2 was from BD Biosciences (San Jose, CA). Lysis buffer and BCA protein assay were from Beyotime (Shanghai, China).
3
Hippocampal Protein Extraction and Western Blotting
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The rat hippocampi were homogenized and placed in a radioimmunoprecipitation assay lysis buffer containing protease and phosphatase inhibitors and phenylmethylsulfonyl fluoride. The samples were lysed on ice for 30 minutes and centrifuged at 13,000 r/min for 10 minutes at 4°C, and the supernatant was aspirated. The bicinchoninic acid assay was performed to determine the protein concentration. The samples were then stored at −80°C until further use. Western blotting was performed using standard protocols [33 (link)]. The membranes were incubated overnight with primary antibodies, including anti-NLRP3 (1:500, Novus, USA), anti-GSDMD (1:1000, Abcam, UK), anti-caspase-1 (1:1000, Novus, USA), and anti-caspase-11 (1:200, Novus, USA), and then with the appropriate secondary antibodies. ImageJ software was used to measure the gray values of the target bands.
4
Testicular Protein Expression Analysis
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Testicular tissues were collected from mice 36 h after UPEC injection and homogenized in radioimmunoprecipitation assay buffer containing a protease inhibitor cocktail (Sigma, P8340), followed by incubation on a shaker at 4 °C for 1 h. The lysates were centrifuged at 12,000×g for 15 min at 4 °C, and then the supernatants were collected to assay protein concentrations using the bicinchoninic acid protein assay. Equal quantities of the samples were resolved using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred onto nitrocellulose membranes, and incubated with the indicated antibodies. The antibodies and working dilutions are listed as follows: anti-NLRP3 (1:1000, Adipogen, AG-20B-0014), anti-ASC (1:1000, Santa Cruz, SC-22514-R), anti- CASPASE-1 (1:1000, Adipogen, AG-20B-0042), anti- CASPASE-11 (1:1000, Novus, NB120-10454), anti-GSDMD (1:1000, Abcam, ab209845), β-actin (1:1000, Sigma, A1978), IRDye 680RD anti-mouse (1:5000, LI-COR Biosciences, 926-68070), IRDye 800CW anti-rabbit (1:5000, LI-COR Biosciences, 926-32211), and goat anti-rabbit-HRP (1:3000, Thermo Fisher, 31402).
5
Quantification of Cell Death Pathways
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To analyze cell death pathways induced in ESCRT-III-silenced cells, total (live and dead) supernatant and adherent cells were collected and counted. 1 × 106 cells were pelleted and lysed in 100 μL 2XSB buffer without DTT (4% SDS (#EU0660, Euromedex), 0.12 M Tris pH 6,8 (#10708976001, Sigma-Aldrich), glycerol 10% (#24388.295, VWR), dash of bromophenol blue (#B-5525, Sigma-Aldrich)) (1 million cells lysed per 100 μL buffer). When needed, DTT (#P2325, ThermoFisher) was added extemporaneously at 100 mM final concentration. Samples were immediately boiled for 5 min at 95°C, quickly spun and frozen at −20°C until use. The following antibodies were used for immunoblotting detection: anti-caspase-3 (#9662, Cell Signaling Technologies), anti-caspase-8 (#9429, Cell Signaling Technologies), anti-caspase-1 (produced in house – Peter Vandenabeele’s lab and Schotte et al. (2004) (link), anti-caspase-11 (#120–10454, Novus Biologicals), anti-cleaved Parp (#9544, Cell Signaling Technologies), anti-Gasdermin-D (Genentech), anti-IL-1β (#GTX74034, Genetex), anti MLKL (#MABC604, Millipore (reducing conditions) or #SAB1302339, Sigma-Aldrich (non-reducing conditions)), anti-phosphoMLKL (#ab196436, Abcam), anti-RIPK3 (#R4277, Sigma-Aldrich), anti-RIPK1 (#610459, BD Biosciences).
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