Heart tissue lysates (10 μg) were boiled in the Laemmli buffer for 5 minutes, and loaded into 10–20% ready gels (Biorad). Protein bands were resolved by SDS-PAGE at 100 V. Electrotransferring was performed at 100 V for 30 minutes onto PVDF membranes. After washing, PVDF was resolved proteins were blocked with 5% non-fat milk. These membranes were then used for detection of the proteins of interest. Rabbit anti-mouse TLR4 antibody, rabbit-anti-MyD88 antibody, and goat anti-rabbit IgG antibody were acquired from Abcam (England). The following dilutions of primary antibodies were used: TLR4 (1: 1000), MyD88 (1: 500), NF-κB-p65 (1: 2000), and p-NF-κB-p65 (1: 1000). Primary antibodies were used to incubate the membranes at 4ºC overnight. After TBST washing, the goat anti-rabbit antibodies were added. Chemiluminescent substrates were then added for visualization of protein bands. Expression levels of TLR4, MyD88, and NF-κB were quantified after normalization to the level of β-actin.
Rabbit anti mouse tlr4 antibody
Manufactured by Abcam
Sourced in United Kingdom
Rabbit anti-mouse TLR4 antibody is a primary antibody that specifically binds to the toll-like receptor 4 (TLR4) protein in mouse samples. TLR4 is a pattern recognition receptor that plays a key role in the innate immune response.
Automatically generated - may contain errors
Lab products found in correlation
Ready gel, by Bio-Rad (1 mentions)
Goat anti rabbit igg antibody, by Abcam (1 mentions)
Fitc rat anti mouse cd45, by BioLegend (1 mentions)
Rat anti mouse ly6g pe, by BD (1 mentions)
Fitc rat anti mouse cd11b, by BioLegend (1 mentions)
Palmitic acid, by Merck Group (1 mentions)
Gapdh, by Abcam (1 mentions)
Phospho p38, by Abcam (1 mentions)
Lamin b1, by Abcam (1 mentions)
β actin, by Abcam (1 mentions)
Pe rat anti mouse f4 80, by BioLegend (1 mentions)
Phospho iκbα, by Abcam (1 mentions)
Phospho p65, by Abcam (1 mentions)
Tak 242, by MedChemExpress (1 mentions)
2 protocols using rabbit anti mouse tlr4 antibody
1
Western Blot Analysis of TLR4, MyD88, NF-κB
2
Inflammatory Signaling Pathway Characterization
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The kits for alanine aminotransferase (ALT), aspartate aminotransferase (AST), and triglyceride (TG) were supplied by Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Enzyme-linked immunosorbent assay (ELISA) kits for TNF-α and IL-1β were purchased from Bender MedSystems (Vienna, Austria). PE rat anti-mouse Ly6G was obtained from BD Biosciences (New Jersey, USA). Neutrophils and F4/80 antibodies were obtained from Thermo Scientific (Rockford, IL, USA). Rabbit anti-mouse TLR4 antibody, rabbit anti-mouse phospho-IRAK1, phospho-p38, phosphor-IKKβ, IKKβ, phospho-IκBα, IκBα, phospho-p65, p38, p65, β-actin, Lamin B1, and GAPDH were purchased from Abcam (Cambridge, UK). PE rat anti-mouse F4/80, FITC rat anti-mouse CD11b, and FITC rat anti-mouse CD45 antibodies were from Biolegend, Inc. (San Diego, USA). TLR4-specific antagonist TAK-242 was from MedChemExpress LLC (Shanghai, China). Palmitic acid (PA) was from Sigma-Aldrich (St. Louis, USA).
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