Samples were lysed in RIPA buffer lysis containing protease inhibitors (PMSF) (Beyotime, China) at 4 °C, and used bicinchoninic acid (BCA) assay to determine the concentration. To further dilute the sample, 6× SDS-PAGE loading buffer was added and boiled (100 °C) for 10 min. An equal amount of the samples (20 μg) was acceded, electrophoresed on a 10% SDS-polyacrylamide denaturing gel, and then transferred to a polyvinylidene fluoride (PVDF) membrane. Anti-HMGB1 (1:1000; Servicebio, Wuhan, China), anti-p65 (1:1000; Bioss, Beijing, China), anti-p-p65 (1:1000; Sigma, Ronkonkoma, NY, USA), anti-TLR4 (1:1000; Sigma, USA), anti-MYD88, anti-NLRP3, anti-IL-1β, anti-pro-caspase-1 (1:500; Wanleibio, Shenyang, China), anti-ASC (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-GAPDH (1:5000; Bioss, China) as the primary antibody. With GAPDH as a reference, and the protein was measured and analyzed by Image J 1.48V and Image Lab 4.0 Software (Bio-Rad, Hercules, CA, USA).
Anti tlr4
Manufactured by Merck Group
Sourced in United States
Anti-TLR4 is a lab equipment product that functions as an antagonist for Toll-like receptor 4 (TLR4). TLR4 is an important innate immune receptor that recognizes pathogen-associated molecular patterns. The Anti-TLR4 product can be used in research applications to study the role of TLR4 signaling in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Image lab 4, by Bio-Rad (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions)
Anti gapdh, by Bioss Antibodies (1 mentions)
Anti hmgb1, by Wuhan Servicebio Technology (1 mentions)
Gapdh, by Bio-Rad (1 mentions)
Anti p65, by Bioss Antibodies (1 mentions)
Anti p p65, by Merck Group (1 mentions)
Anti nlrp3, by Wanlei (1 mentions)
Anti il 1β, by Wanlei (1 mentions)
Anti pro caspase 1, by Wanlei (1 mentions)
Anti asc, by Santa Cruz Biotechnology (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Anti α sma, by Merck Group (1 mentions)
Anti tsp1, by Novus Biologicals (1 mentions)
2 protocols using anti tlr4
1
Western Blot Analysis of HMGB1 and NF-κB Pathway
2
Protein Expression Analysis via Immunoblotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total protein was extracted from frozen liver tissue or cultured cells. From each sample, 30 μg protein was separated in a 10% SDS-polyacrylamide gel electrophoresis and transferred onto a nitrocellulose membrane. Protein expression was determined by immunoblotting with the following antibodies: anti-TSP1 (Novus Biologicals), anti-α-SMA (Sigma), anti-SMPDL3B (Proteintech, IL, USA), anti-Toll-like receptor 4 (anti-TLR4) (Sigma), and anti-β-actin (Sigma). Membranes were blocked and then incubated with primary antibodies, followed by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies. Labelled proteins were detected with an enhanced chemiluminescence system (Pierce).
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