Proteins were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE ) and transferred to polyvinylidene fluoride (PVDF ) membranes. After blocking with phosphate-buffered saline with Tween 20 (PBST) containing 5% fat-free milk, PVDF membranes were coincubated with the antibodies anti-NRF1(1: 1,000, Proteintech, Wuhan, China); anti-NRF2(1: 1,000, Proteintech); anti-PCNA(1: 1,000, Proteintech); anti-β-actin (1: 1,000, Sigma-Aldrich, St. Louis, MO), anti-IκBα (1: 1,000, Proteintech, China), anti-P50 (1: 1,000, Proteintech), anti-NLRP3 (1: 1,000, Wanleibio, Shenyang, China), anti-ASC (1: 1,000, Wanleibio), anti-pro-Caspase1(1: 1,000, Wanleibio); anti-Caspase1-p20 (1: 1,000, Wanleibio) at 4°C overnight. Subsequently, PVDF membranes were incubated with the corresponding secondary antibody at 37°C for 1 h. Proteins were detected using the ECL kit (Beyotime, Beijing), and visualized using a Tanon-5200Multi (Tanon, Shanghai, China) device. Densitometry analysis was performed using the Image J software (National Institutes of Health, Bethesda, MD).
Anti caspase1 p20
Manufactured by Wanlei
Sourced in China
Anti-Caspase1-p20 is a laboratory equipment used to detect the presence and activity of Caspase-1, a protease enzyme involved in the inflammatory response. This product provides a reliable method for measuring Caspase-1 levels in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti pro caspase 1, by Wanlei (2 mentions)
Anti asc, by Wanlei (2 mentions)
Anti β actin, by Merck Group (1 mentions)
Ecl kit, by Beyotime (1 mentions)
Anti pcna, by Proteintech (1 mentions)
Anti nlrp3, by Wanlei (1 mentions)
Anti nrf1, by Proteintech (1 mentions)
Anti iκbα, by Proteintech (1 mentions)
Anti nrf2, by Proteintech (1 mentions)
Anti p nf κb p65, by Cell Signaling Technology (1 mentions)
Anti nf κb p65, by Cell Signaling Technology (1 mentions)
Ecl detection system, by Thermo Fisher Scientific (1 mentions)
Anti gr, by Bioss Antibodies (1 mentions)
Anti il 18, by Wanlei (1 mentions)
Anti lamin b, by Wanlei (1 mentions)
Anti β actin, by Bioss Antibodies (1 mentions)
Anti il 1β, by Wanlei (1 mentions)
Anti nlrp3, by Bioss Antibodies (1 mentions)
Anti iba1, by Abcam (1 mentions)
Nuclear and cytoplasmic protein extraction kit, by Applygen (1 mentions)
2 protocols using anti caspase1 p20
1
Western Blot Analysis of NRF, PCNA, and Inflammatory Markers
2
Western Blot Analysis of NF-κB Activation
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Equal amounts of protein samples (28 μg) were subjected to a SDS-PAGE and transferred to PVDF membranes. Membranes were blocked in 5% milk in TBST for 2 h followed by incubation with the following primary antibodies as follows: anti-p-NF-κB P65 (1:1000, Cell Signaling Technology), anti-NF-κB P65 (1:1000, Cell Signaling Technology), anti-Iba-1 (1:1000, Abcam), anti-β-actin (1:7500, Bioss antibodies), anti-GR (1:1000, Bioss antibodies), anti-NLRP3 (1:1500), anti-ASC (1:500), anti-pro-caspase 1 (1:500), anti-caspase-1 p20 (1:750), anti-pro-IL-1β (1:10000), anti-IL-1β (1:750), anti-IL-18 (1:1000) and anti-Lamin B (1:750) both from Wanlei Biotechnology (Shenyang, China). After three times washing with TBST, the membranes were incubated with appropriate secondary antibody. The protein bands were visualized by the ECL detection system (Thermo Scientific, Waltham, MA, United States) and quantified using Image J software. Nuclear and Cytoplasmic Protein Extraction Kit (Applygen Technologies, Co., Ltd., Beijing, China) was used to extract the cytoplasmic/nuclear proteins of NF-κB P65 according to the manufacturer’s protocol.
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