mouse anti human tlr4 mabs hta125  (Bio-Rad)

Journal: BMC veterinary research

Article Title: Escherichia coli HPI-induced duodenitis through ubiquitin regulation of the TLR4/NF-κB pathway.

doi: 10.1186/s12917-025-04515-3

Figure Lengend Snippet: Fig. 1 E. coli HPI promotes the expression of Ub, TLR4, and NF-κB. A The effects of TLR4 and NF-κB on IPEC-J2 cells treated with E. coli HPI for 6 and 12 h were observed by immunofluorescence (scale bar, 10 μm). B Relative mean fluorescence of TLR4 at 6 and 12 h after E. coli HPI infection. Randomly selected 3 fields of view. C Nuclear NF-κB p65 expression levels at 6 and 12 h after E. coli HPI infection. Randomly selected 3 fields of view. D Ub mRNA levels at 6 and 12 h after E. coli HPI infection (n = 3). All data are shown as the mean ± SD. *** p < 0.001, and ** p < 0.01, and * p < 0.05

Article Snippet: After serum blocking, the sections were incubated overnight at 4 °C with primary antibodies against TLR4 (#14358) and NF-κB (#6956S) from CST (Beijing, China), TNF-α (#52B83), and IL-1β (#ab2105) from abcam (Shanghai, China).

Techniques: Expressing, Immunofluorescence, Fluorescence, Infection

Journal: BMC veterinary research

Article Title: Escherichia coli HPI-induced duodenitis through ubiquitin regulation of the TLR4/NF-κB pathway.

doi: 10.1186/s12917-025-04515-3

Figure Lengend Snippet: Fig. 2 The TLR4/NF-kB pathway is involved in E. coli infection: A functional enrichment: the enriched KEGG signaling pathways were selected to dem onstrate the primary biological actions of major potential mRNA; B volcano plot: the volcano plot was constructed using the fold change values and p-adjusted values. Red dots represent genes with significant fold change value and p value, gray dots represent genes with insignificant fold change value and p value; C the heatmap of the differential gene expression, with different colors representing the trend of gene expression in different groups; D functional enrichment: the GO functional enrichment was selected to demonstrate the primary biological actions of major potential mRNA; E network of interacting proteins predicted via TLR4

Article Snippet: After serum blocking, the sections were incubated overnight at 4 °C with primary antibodies against TLR4 (#14358) and NF-κB (#6956S) from CST (Beijing, China), TNF-α (#52B83), and IL-1β (#ab2105) from abcam (Shanghai, China).

Techniques: Infection, Functional Assay, Protein-Protein interactions, Construct, Gene Expression

Journal: BMC veterinary research

Article Title: Escherichia coli HPI-induced duodenitis through ubiquitin regulation of the TLR4/NF-κB pathway.

doi: 10.1186/s12917-025-04515-3

Figure Lengend Snippet: Fig. 5 E. coli HPI regulates TLR4/NF-κB pathway by upregulating Ub. A IHC was performed to measure the expression of TLR4, TNF-α, IL-1β, and NF-κB p65 in duodenal tissues. The images of TLR4, IL-1β, and TNF-α were captured at 200× magnification (scale bar, 50 μm), while the image of NF-κB p65 was captured at 1000× magnification (scale bar, 10 μm). B The protein-positive area of TLR4, NF-κB p65, TNF-α, and IL-1β was quantified. Randomly selected 3 fields of view. C The mRNA levels of TLR4, Myd88, IκB-α, and NF-κB were measured in BALB/c mice infected with E. coli HPI (n = 3). All data are shown as the mean ± SD. *** p < 0.001, and ** p < 0.01, and * p < 0.05

Article Snippet: After serum blocking, the sections were incubated overnight at 4 °C with primary antibodies against TLR4 (#14358) and NF-κB (#6956S) from CST (Beijing, China), TNF-α (#52B83), and IL-1β (#ab2105) from abcam (Shanghai, China).

Techniques: Expressing, Infection

Journal: BMC veterinary research

Article Title: Escherichia coli HPI-induced duodenitis through ubiquitin regulation of the TLR4/NF-κB pathway.

doi: 10.1186/s12917-025-04515-3

Figure Lengend Snippet: Fig. 6 Schematic diagram illustrating E. coli HPI-induced duodenitis through ubiquitin regulation of the TLR4/NF-κB pathway

Article Snippet: After serum blocking, the sections were incubated overnight at 4 °C with primary antibodies against TLR4 (#14358) and NF-κB (#6956S) from CST (Beijing, China), TNF-α (#52B83), and IL-1β (#ab2105) from abcam (Shanghai, China).

Techniques: Ubiquitin Proteomics

Journal: Critical Care

Article Title: Toll-like receptor 4 in phagocytosis of Escherichia coli by endotoxin-activated human neutrophils in whole blood

doi: 10.1186/cc11767

Figure Lengend Snippet: Influence of antibodies against TLR4, CD14 and CD11b on neutrophil phagocytic activity in whole blood . Control cells without any incentives (C), cells activated by S-LPS (C1) and cells activated by Re-LPS (C2).

Article Snippet: Fluorescein-labeled bioparticles E. coli K12 (Molecular Probes), mouse anti-human TLR4 mAbs HTA125 (IgG2a isotype; Serotec), anti-human CD14 clone UCHM-1 mAbs (IgG2a isotype; Sigma), and anti-human CD11b mAbs clone 44 (IgG1 isotype; Sigma).

Techniques: Activity Assay

Journal: Frontiers in Pharmacology

Article Title: Ameliorative effect of Berberidis radix polysaccharide selenium nanoparticles against carbon tetrachloride induced oxidative stress and inflammation

doi: 10.3389/fphar.2022.1058480

Figure Lengend Snippet: (A) BRP-SeNPs protected mice from CCl 4 -induced liver injury via Nrf2/Keap1/MKP1/JNK and TLR4/MAPK signaling pathways. (B) Quantification of (A) Nrf2, (B) Keap1, (C) MKP1, (D) p-JNK, (E) p-ASK1, (F) p-MKK4, (G) TLR4, (H) p-p38, (I) p-ERK expression in cytoplasm. The different letters indicate significant differences ( p < 0.05).

Article Snippet: Antibodies of mouse TLR4, p-JNK, p-ERK, p-p38, p-ASK, p-MKK4, Keap1, Nrf2 were purchased from Cell Signaling Technology (Beverly, MA, United States).

Techniques: Protein-Protein interactions, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Endoplasmic Reticulum Homeostasis Regulates TLR4 Expression and Signaling in Mast Cells

doi: 10.3390/ijms231911826

Figure Lengend Snippet: TLR4 surface expression and function is supported by the IRE1/XBP1 pathway. TNF and IL-6 levels in the supernatants in response to LPS stimulation (1 μg/mL, 4 h) were measured by ELISA for an equal number of YFP-sorted XBP1 KO and DKO PMCs, as compared to WT PMCs. The average of three independent experiments ± SD is shown for ( A ) IL-6 and ( B ) TNF. * p < 0.05. ( C ) Flow cytometry analysis of TLR4 cell surface expression of XBP1 KO (green histogram) and DKO PMCs (red histogram), as compared to WT PMCs (blue histogram). ( D ) Quantification of TLR4 expression levels in WT, XBP1 KO, and DKO PMCs. Data are presented as the average ± SD of the mean fluorescence intensity minus the background (ΔMFI) of three independent analyses. * p < 0.05. ( E ) Flow cytometry analysis of TLR2 expression in WT, XBP1 KO, and DKO PMCs (blue, green, and red histograms, respectively). ( F ) RT-qPCR analysis of sXBP1 in PMCs treated with DMSO or 60 μM STF, 1 h prior to stimulation with LPS (1 μg/mL) for 3, 6, and 12 h. Bars are represented as the mean of the relative mRNA levels normalized to GAPDH ± SD of three independent experiments. Cytokines levels in the supernatants for ( G ) IL-6 and ( H ) TNF were determined by ELISA. Each bar represents the mean ± SD of three independent experiments. * p < 0.05.

Article Snippet: TLR4 mAb mouse (sc-293072) was obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Fluorescence, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Endoplasmic Reticulum Homeostasis Regulates TLR4 Expression and Signaling in Mast Cells

doi: 10.3390/ijms231911826

Figure Lengend Snippet: IRE1 activation supports cooperativity between TLR4 and FcεRI in CBMCs. ( A ) IgE-sensitized CBMCs were pretreated with DMSO or STF (30 or 60 μM) for 1 h followed by activation with rabbit anti-human IgE Ab (5 μg/mL, 4 h). Equal amounts of whole cell protein extracts were analyzed by immunoblotting for TLR4 and p97 as a loading control. RT-qPCR analysis of sensitized CBMCs after activation with rabbit anti-human IgE Ab (5 μg/mL), stimulation with 100 ng/mL LPS, or both treatments for 4 h, for ( B ) sXBP1, ( C ) TNF, and ( D ) IL-6. Bars represent the mean of relative mRNA levels normalized to β-actin ± SD of three independent experiments. * p < 0.05. ( E ) IgE-sensitized CBMCs were pretreated with DMSO or STF (60 μM) for 1 h followed by activation with rabbit anti-human IgE Ab (5 μg/mL), 100 ng/mL LPS, or both for 4 h. IL-6 levels in the supernatants were quantified by ELISA. Bars represent the mean ± SD of three independent experiments. * p < 0.05. PMCs were sensitized overnight with mouse IgE-anti-DNP (0.5 µg/mL, 18 h). Later, cells were stimulated with 100 ng/mL LPS, activated with DNP-BSA (25 or 50 ng/mL), or stimulated with both for 4 h. Levels of IL-6 ( F ) and TNF ( G ) in supernatants were determined by ELISA. Each bar represents the mean ± SD of three independent experiments. * p < 0.05.

Article Snippet: TLR4 mAb mouse (sc-293072) was obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Activation Assay, Western Blot, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Endoplasmic Reticulum Homeostasis Regulates TLR4 Expression and Signaling in Mast Cells

doi: 10.3390/ijms231911826

Figure Lengend Snippet: List of murine primer sequences for RT-qPCR.

Article Snippet: TLR4 mAb mouse (sc-293072) was obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques:

Journal: Cell reports

Article Title: Candida albicans oscillating UME6 expression during intestinal colonization primes systemic Th17 protective immunity

doi: 10.1016/j.celrep.2022.110837

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-mouse TLR4/MD2 (clone MTS510) , Hycult Biotech , Cat#HM1029; RRID:AB_533218.

Techniques: Recombinant, Staining, Conjugation Assay, SYBR Green Assay, Software

Journal: Journal of Healthcare Engineering

Article Title: Butylphthalide Inhibits TLR4/NF- κ B Pathway by Upregulation of miR-21 to Have the Neuroprotective Effect

doi: 10.1155/2022/4687349

Figure Lengend Snippet: Effects of Butylphthalide treatment and miR-21 overexpression on TLR4/NF- κ B of Neuro2A cells. (a) The protein expression levels of p-TLR4, p-TLR4, p-NF- κ B, and NF- κ B in OGD/R + Butylphthalide group, OGD/R + miR-21 mimics group, OGD/R + miR-NC group, and control group. (b) Statistical results showed that the proportion of p-TLR4/TLR4 in OGD/R + Butylphthalide group and OGD/R + miR-21 mimics group was significantly lower than that in OGD/R + miR-NC group, and the proportion of p-TLR4/TLR4x in OGD/R + miR-NC group was higher than that in control group. (c) Statistical results showed that the proportion of p-NF- κ B/NF- κ B in OGD/R + Butylphthalide group and OGD/R + miR-21 mimics group was significantly lower than that in OGD/R + miR-NC group, and the proportion of p-NF- κ B/NF- κ B in OGD/R + miR-NC group was higher than that in control group. ∗ P < 0.05, ∗ ∗ P < 0.01, ∗ ∗ ∗ P < 0.001, ∗ ∗ ∗ ∗ P < 0.0001.

Article Snippet: The membrane was sealed in TBST buffer containing 5% skim milk for 1 hour and then incubated overnight at 4°C with the following primary antibodies: anti-mouse-p-tlr4 (Santa Cruz, 1 : 1000), anti-mouse TLR4 (CST, 1 : 1000), anti-mouse-p-nf- κ B (Santa Cruz, 1 : 1000), anti-mouse-NF- κ B (Santa Cruz, 1 : 1000), anti-mouse-casepse3 (abcam,1 : 1000), anti-mouse-Bax (abcam,1 : 1000), anti-mouse-Bcl-2 (abcam,1 : 1000), and anti-mouse-GAPDH (Santa Cruz, 1 : 1000).

Techniques: Over Expression, Expressing, Control

Journal: iScience

Article Title: Stabilin receptors clear LPS and control systemic inflammation

doi: 10.1016/j.isci.2021.103337

Figure Lengend Snippet: Clearance of LPS by LSEC is via a receptor-mediated mechanism; however, TLR4 is not involved in the rapid clearance of LPS by LSEC (A) Plot for 488-LPS bound to LSEC in a dose-dependent manner versus concentration of 488-LPS along with excess of unlabeled LPS, HDL, and LBP incubated at 37°C for 1 h. (B) The curve plots the remainder of 3 H/ 14 C double-labeled LPS in blood circulation versus time in WT C57BL/6 and TLR4-KO mice at various time points after intravenous (IV) infusion. Each data point represents the mean and SD of data from three mice/biological replicates. The figures represent two different experiments.

Article Snippet: The anti-mouse TLR4 antibody (Cell Signaling USA) and mouse anti-GAPDH (Santa Cruz) were used at a working concentration of 10μg/ml.

Techniques: Concentration Assay, Incubation, Labeling

Journal: iScience

Article Title: Stabilin receptors clear LPS and control systemic inflammation

doi: 10.1016/j.isci.2021.103337

Figure Lengend Snippet: TLR4 is weakly expressed in the liver compared with all major internal organs, but LSEC and KC express TLR4 similar to professional macrophages (A) An ECL-developed immunoblot using rabbit anti-mouse TLR4 Ab showing TLR4 expression in spleen lysates of WT C57BL/6 and TLR4-KO mouse, RAW 264.7 and HEK cell line lysates prepared as described in materials and methods. (B) Reprobe of A showing ECL-developed immunoblot of mouse anti-GAPDH antibody as a loading control. (C) An ECL-developed immunoblot using rabbit anti-mouse TLR4 Ab showing TLR4 expression in major organ lysates of BALB/c mice and standard RAW cell lysates. (D) Bar graph expressing the means and SD of TLR4 expression distributed to each organ, shown in C, after factoring total weight of the organ. (E) An ECL-developed immunoblot using rabbit anti-mouse TLR4 Ab showing TLR4 expression in major organ lysates of C57BL/6 mice and standard RAW cell lysates. (F) Bar graph expressing the means and SD of TLR4 expression distributed to each organ, shown in E after factoring total weight of the organ. (G) An ECL-developed immunoblot using rabbit anti-mouse TLR4 Ab showing TLR4 expression in different liver cell lysates of C57BL/6 mice. (H) A reprobe of G showing the expression of GAPDH. The numbers depicted represent the MW for each respective band in kDa. Each figure is a representative image from three mice, and the bar graph compiles data from three mice. Values of all significant correlations are given with degree of significance manner versus concentration of 488-LPS along with indicated ∗p < 0.05.

Article Snippet: The anti-mouse TLR4 antibody (Cell Signaling USA) and mouse anti-GAPDH (Santa Cruz) were used at a working concentration of 10μg/ml.

Techniques: Western Blot, Expressing, Control, Concentration Assay

Journal: iScience

Article Title: Stabilin receptors clear LPS and control systemic inflammation

doi: 10.1016/j.isci.2021.103337

Figure Lengend Snippet: LSEC produce both pro-inflammatory and anti-inflammatory cytokines in response to TLR4 (A) Volcano plot showing expression of differentially expressed transcript of purified mouse LSEC that were stimulated with LPS or left untreated for 6 h and the cell free culture supernatants were harvested and the cells were lysed in TRizol. Total RNA was extracted, and the mRNA samples were analyzed using NanoString assay on the mouse Pan cancer Immune panel, followed by data analysis in nSolver software. (B) The amount of TNF-α and IL10 cytokine levels in the supernatant were determined by ELISA. (C) The graphs shown are the fold change of TNF-α, IL-1β, IL-1α, IL-15, IL-6, and IL10 mRNA of untreated samples. Values of all significant correlations are given with degree of significance indicated. ∗p < 0.005.

Article Snippet: The anti-mouse TLR4 antibody (Cell Signaling USA) and mouse anti-GAPDH (Santa Cruz) were used at a working concentration of 10μg/ml.

Techniques: Expressing, Purification, Software, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Stabilin receptors clear LPS and control systemic inflammation

doi: 10.1016/j.isci.2021.103337

Figure Lengend Snippet:

Article Snippet: The anti-mouse TLR4 antibody (Cell Signaling USA) and mouse anti-GAPDH (Santa Cruz) were used at a working concentration of 10μg/ml.

Techniques: Virus, Labeling, Clinical Proteomics, Recombinant, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Gene Expression, Stable Transfection, Expressing, Modification, Knock-Out, Software