rabbit polyclonal antibody to tlr4  (Affinity Biosciences)

Journal: Scientific Reports

Article Title: In vivo mechanisms of resveratrol liposomes in targeting the TLR4/NLRP3 pathway in lung cancer

doi: 10.1038/s41598-025-14587-8

Figure Lengend Snippet: Resveratrol liposome downregulates the TLR4 pathway in lung cancer. ( A ) Representative images of TLR4 and MyD88 immunofluorescence in tumor tissues (scale bar: 50 μm); ( B-C ) Fluorescence intensity of TLR4 and MyD88 in tumor tissues; ( D ) mRNA relative expression of the TLR4/MyD88 pathway in tumor tissues; ( E-F ) Protein expressions of TLR4, MyD88, IRAK4, and p-TAK1 in tumor tissues measured by Western blotting; original western blot scans are shown in Supplementary Fig. 10 Data are presented as mean ± standard deviation; (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns indicates no statistically significant difference between samples).

Article Snippet: Primary antibodies used included: NLRP3 antibody (Hangzhou Huaan, ET1610-93), Caspase-1 p20 antibody (Hangzhou Huaan, ER1905-47), IRAK4 antibody (Hangzhou Huaan, EM1709-86), TLR4 polyclonal antibody (Abclonal, A5258), MyD88 polyclonal antibody (Solarbio, K001785P), ASC antibody (Cell Signaling Technology, #67824), TAK1 (phospho S439) antibody (Abcam, EPR2863), and GADPH (Servicebio, GB15002).

Techniques: Immunofluorescence, Fluorescence, Expressing, Western Blot, Standard Deviation

Journal: Scientific Reports

Article Title: In vivo mechanisms of resveratrol liposomes in targeting the TLR4/NLRP3 pathway in lung cancer

doi: 10.1038/s41598-025-14587-8

Figure Lengend Snippet: Resveratrol liposome downregulated the NLRP3/ASC/Caspase-1 pathway. ( A ) qPCR analysis of mRNA relative expression levels of NLRP3, Prcard, caspase-1, and GSDMD in tumor tissues; ( B ) Representative images of immunofluorescence staining for TLR4 and MyD88 in tumor tissues (scale bar: 50 μm); ( C-D ) Protein expressions of NLRP3, ASC, Casp-1, Casp-1(P20) in tumor tissues measured by Western blotting; original western blot scans are shown in Supplementary Fig. 11. Data are presented as mean ± standard deviation (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns indicates no statistically significant difference between samples).

Article Snippet: Primary antibodies used included: NLRP3 antibody (Hangzhou Huaan, ET1610-93), Caspase-1 p20 antibody (Hangzhou Huaan, ER1905-47), IRAK4 antibody (Hangzhou Huaan, EM1709-86), TLR4 polyclonal antibody (Abclonal, A5258), MyD88 polyclonal antibody (Solarbio, K001785P), ASC antibody (Cell Signaling Technology, #67824), TAK1 (phospho S439) antibody (Abcam, EPR2863), and GADPH (Servicebio, GB15002).

Techniques: Expressing, Immunofluorescence, Staining, Western Blot, Standard Deviation

Journal: Nature Communications

Article Title: RIPK1 kinase drove brain microvascular endothelial cells death and blood-brain barrier disruption in neonatal Escherichia coli meningitis

doi: 10.1038/s41467-025-62760-4

Figure Lengend Snippet: a Co-IP of pro-caspase-8 with RIPK1, RIPK3, ZBP1, and ASC in WT, RIPK1 KO, or ZBP1 KO cells infected with RS218 (MOI = 1). b Co-IP of pro-caspase-8 with RIPK1, RIPK3, ZBP1, and ASC in hBMECs expressing RIPK1 WT or D138N under RS218 infection (MOI = 1). c Immunoblot of RIPK1 total protein and phosphorylation levels at S161 and S166 in WT, TNFR1 KO, TLR4 KO, or double KO cells post RS218 infection (MOI = 1). Blots for GAPDH served as loading controls. d Co-IP of pro-caspase-8 with RIPK1, RIPK3, ZBP1, and ASC in hBMECs expressing RIPK1 WT, S161N, or S166N cells (MOI = 1). e , f In WT, TNFR1 KO, TLR4 KO, or double KO cells, the cytotoxicity was assessed by LDH release ( e , n = 3 biologically independent samples), and cell death kinetics were measured by PI uptake ( f , n = 3 biologically independent samples). g Co-IP analysis of RIPK1 with TNFR1 and TLR4 interactions in WT and ZBP1 KO cells. h Co-IP of pro-caspase-8 with RIPK1, RIPK3, ZBP1, and ASC in WT, TNFR1 KO, TLR4 KO or double KO cells (MOI = 1). i Immunoblot of RIPK1 total protein and phosphorylation levels at S161 in WT or ZBP1 KO cells. Blots for GAPDH served as loading controls. All experiments were representative of at least three independent experiments with similar results. Bars indicated the mean plus SD. Statistical significance was determined by one-way ANOVA with Tukey’s post-hoc test, with P- values denoted as follows: **** P < 0.0001, *** P < 0.0005, ** P < 0.005, * P < 0.05, n.s., no significant difference. Exact P- values were provided in the source data. Source data are provided as a Source Data file.

Article Snippet: The PVDF membranes then blotted with rabbit anti-caspase 3/p17 antibody (Proteintech, 19677-1-AP, 1:2000), mouse anti-PARP1 Monoclonal antibody (Proteintech, 66520-1-Ig, 1:20000), rabbit anti-GSDMD-NT polyclonal antibody (Bioworld, BS67358, 1:2000), rabbit anti-MLKL Antibody (ABclonal, A19685, 1:2000), rabbit anti-Phospho-MLKL (Ser358) Antibody (Affinity, AF7420, 1:2000), rabbit anti-RIPK1-Specific Polyclonal antibody (ABclonal, A19580, 1:2000), rabbit anti-Phospho-RIPK1 (Ser161) Antibody (Abmart, TA7377, 1:2000), rabbit anti-Phospho-RIPK1 (Ser166) Antibody (Abmart, TA2398S, 1:2000), rabbit anti-RIP3 Polyclonal antibody (Proteintech, 17563-1-AP, 1:2000), rabbit anti-RIP3 (phospho S227) antibody (abcam, ab209384, 1:2000), mouse anti-caspase-1/p10 antibody (Santa Cruz Biotechnology, sc-56036, 1:500), rabbit anti-caspase 8/p43/p18 Polyclonal antibody (Proteintech, 13423-1-AP, 1:1000), rabbit anti-ASC/TMS1 Polyclonal antibody (Proteintech, 10500-1-AP, 1:20000) or rabbit anti-TRIF (TICAM1) antibody (abcam, ab302562, 1:1000) or rabbit anti-ZBP1 Polyclonal antibody (Proteintech, 13285-1-AP, 1:5000) or rabbit anti-TLR4 Polyclonal antibody (Proteintech, 19811-1-AP, 1:2000) or rabbit anti-TNF Receptor I antibody (abcam, ab223352, 1:1000) or rabbit anti-GAPDH polyclonal antibody (Bioworld, AP0063, 1:10000), followed by incubation overnight at 4 °C.

Techniques: Co-Immunoprecipitation Assay, Infection, Expressing, Western Blot, Phospho-proteomics

Journal: Nature Communications

Article Title: RIPK1 kinase drove brain microvascular endothelial cells death and blood-brain barrier disruption in neonatal Escherichia coli meningitis

doi: 10.1038/s41467-025-62760-4

Figure Lengend Snippet: a , b Immunoblot analysis of cleaved caspase-3, PARP1, GSDMD, and phosphorylated RIPK1, RIPK3, and MLKL in RIPK1 WT and RIPK1 S161N cells ( a ) or in WT, RIPK1 KO, and RIPK3 KO cells ( b ) after RS218 infection (MOI = 1). Blots for GAPDH served as loading controls. c Immunoblot analysis of total and phosphorylated levels of RIPK1, RIPK3, and MLKL in WT, TNFR1 KO, TLR4 KO, or double KO hBMECs expressing RIPK1 WT or S161N. Blots for GAPDH served as loading controls. d TLR4 was co-immunoprecipitated with RIPK3 and TRIF in RIPK1 WT and S161N cells. e In RIPK1 S161N background, TLR4 was co-immunoprecipitated with RIPK3 and TRIF in TRIF WT, TRIF KO, and TRIF mRHIM cells. f In the RIPK1 S161N background, immunoblot analysis of total and phosphorylated levels of RIPK1, RIPK3, and MLKL in TRIF WT, TRIF KO, and TRIF mRHIM cells. Blots for GAPDH served as loading controls. All experiments were representative of at least three independent experiments with similar results. Source data are provided as a Source Data file.

Article Snippet: The PVDF membranes then blotted with rabbit anti-caspase 3/p17 antibody (Proteintech, 19677-1-AP, 1:2000), mouse anti-PARP1 Monoclonal antibody (Proteintech, 66520-1-Ig, 1:20000), rabbit anti-GSDMD-NT polyclonal antibody (Bioworld, BS67358, 1:2000), rabbit anti-MLKL Antibody (ABclonal, A19685, 1:2000), rabbit anti-Phospho-MLKL (Ser358) Antibody (Affinity, AF7420, 1:2000), rabbit anti-RIPK1-Specific Polyclonal antibody (ABclonal, A19580, 1:2000), rabbit anti-Phospho-RIPK1 (Ser161) Antibody (Abmart, TA7377, 1:2000), rabbit anti-Phospho-RIPK1 (Ser166) Antibody (Abmart, TA2398S, 1:2000), rabbit anti-RIP3 Polyclonal antibody (Proteintech, 17563-1-AP, 1:2000), rabbit anti-RIP3 (phospho S227) antibody (abcam, ab209384, 1:2000), mouse anti-caspase-1/p10 antibody (Santa Cruz Biotechnology, sc-56036, 1:500), rabbit anti-caspase 8/p43/p18 Polyclonal antibody (Proteintech, 13423-1-AP, 1:1000), rabbit anti-ASC/TMS1 Polyclonal antibody (Proteintech, 10500-1-AP, 1:20000) or rabbit anti-TRIF (TICAM1) antibody (abcam, ab302562, 1:1000) or rabbit anti-ZBP1 Polyclonal antibody (Proteintech, 13285-1-AP, 1:5000) or rabbit anti-TLR4 Polyclonal antibody (Proteintech, 19811-1-AP, 1:2000) or rabbit anti-TNF Receptor I antibody (abcam, ab223352, 1:1000) or rabbit anti-GAPDH polyclonal antibody (Bioworld, AP0063, 1:10000), followed by incubation overnight at 4 °C.

Techniques: Western Blot, Infection, Expressing, Immunoprecipitation