rabbit anti il 1r1 antibody  (Bioss)

Journal: Cureus

Article Title: Melittin Alleviates Oxidative Stress Injury in Schwann Cells by Targeting Interleukin-1 Receptor Type 1 to Downregulate Nuclear Factor Kappa B-Mediated Inflammatory Response In Vitro

doi: 10.7759/cureus.65721

Figure Lengend Snippet: (A) and (B): RT-qPCR analysis of reactive mRNA of IL-1β and TNF-α expression in hypoxic SCs and the intervention effect of melittin. (C) Immunofluorescence assays after staining with anti-Fn and anti-IL-1R1 antibodies (scale bars = 50 μm). ****p < 0.0001 versus the control group. Data are presented as mean ± standard deviation. SCs: Schwann cells; RT-qPCR: Quantitative reverse transcription-polymerase chain reaction; IL-1R1: Interleukin-1 receptor type 1; TNF-α: Tumor necrosis factor-alpha; DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: Briefly, cells cultured on coverslips were fixed with 4% paraformaldehyde in PBS for 10 min and then permeabilized with 0.05% Triton X-100 for 10 min. Non-specific binding sites were blocked with 5% goat serum for 1 h. Subsequently, the cells were incubated with a rabbit anti-rat IL-1R1 monoclonal antibody (AF7212, Beyotime, Jiangsu, China) at room temperature for 1.5 h. After washing three times with PBS, the cells were incubated with FITC-conjugated goat anti-rabbit IgG (#7074, Cell Signaling) for 2 h at room temperature.

Techniques: Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Control, Standard Deviation, Reverse Transcription, Polymerase Chain Reaction

Journal: Cureus

Article Title: Melittin Alleviates Oxidative Stress Injury in Schwann Cells by Targeting Interleukin-1 Receptor Type 1 to Downregulate Nuclear Factor Kappa B-Mediated Inflammatory Response In Vitro

doi: 10.7759/cureus.65721

Figure Lengend Snippet: (A) RT-qPCR results showing the expression levels of IL-1R1 and IL-1β. (B) Transmission electron microscope images of ultrastructural alterations in CoCl 2 -induced SCs for each group (Left: ×2500, Right: ×7000), with the red arrows indicating mitochondria. (C) Western blot of C-JUN and GDNF protein expression for quantitative analyses. (D) and (E) Western blot and RT-qPCR results showing the expression levels of VEGF, HIF-1α, IL-1R1, ENO1, AR, SOD, NGF, and iNOS. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 versus control and other groups. Data are presented as mean ± standard deviation. SCs: Schwann cells; GDNF: Glial cell line-derived neurotrophic factor; RT-qPCR: Quantitative reverse transcription-polymerase chain reaction; VEGF: Vascular endothelial growth factor; HIF-1α: Hypoxia-inducible factor 1-alpha; IL-1R1: Interleukin-1 receptor type 1; ENO1: Enolase 1; AR: Aldose reductase; SOD: Superoxide dismutase; NGF: Nerve growth factor; iNOS: Inducible nitric oxide synthase.

Article Snippet: Briefly, cells cultured on coverslips were fixed with 4% paraformaldehyde in PBS for 10 min and then permeabilized with 0.05% Triton X-100 for 10 min. Non-specific binding sites were blocked with 5% goat serum for 1 h. Subsequently, the cells were incubated with a rabbit anti-rat IL-1R1 monoclonal antibody (AF7212, Beyotime, Jiangsu, China) at room temperature for 1.5 h. After washing three times with PBS, the cells were incubated with FITC-conjugated goat anti-rabbit IgG (#7074, Cell Signaling) for 2 h at room temperature.

Techniques: Quantitative RT-PCR, Expressing, Transmission Assay, Microscopy, Western Blot, Control, Standard Deviation, Derivative Assay, Reverse Transcription, Polymerase Chain Reaction

Journal: Cureus

Article Title: Melittin Alleviates Oxidative Stress Injury in Schwann Cells by Targeting Interleukin-1 Receptor Type 1 to Downregulate Nuclear Factor Kappa B-Mediated Inflammatory Response In Vitro

doi: 10.7759/cureus.65721

Figure Lengend Snippet: (A-E) RT-qPCR results showing mRNA expressions of IKK, IKB-α, p65, p60, and IRAK1. (F) Western blot results of protein expression of IKK, IKB-α, p65, p60, and IRAK1. (G-K) Quantitative analyses of IKK, IKB-α, p65, p60, and IRAK1 protein expression. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 versus control and other groups. Data are presented as mean ± standard deviation. RT-qPCR: Quantitative reverse transcription-polymerase chain reaction; IL-1R1: Interleukin-1 receptor type 1; NF-κB: Nuclear factor kappa B.

Article Snippet: Briefly, cells cultured on coverslips were fixed with 4% paraformaldehyde in PBS for 10 min and then permeabilized with 0.05% Triton X-100 for 10 min. Non-specific binding sites were blocked with 5% goat serum for 1 h. Subsequently, the cells were incubated with a rabbit anti-rat IL-1R1 monoclonal antibody (AF7212, Beyotime, Jiangsu, China) at room temperature for 1.5 h. After washing three times with PBS, the cells were incubated with FITC-conjugated goat anti-rabbit IgG (#7074, Cell Signaling) for 2 h at room temperature.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Control, Standard Deviation, Reverse Transcription, Polymerase Chain Reaction

Journal: Cureus

Article Title: Melittin Alleviates Oxidative Stress Injury in Schwann Cells by Targeting Interleukin-1 Receptor Type 1 to Downregulate Nuclear Factor Kappa B-Mediated Inflammatory Response In Vitro

doi: 10.7759/cureus.65721

Figure Lengend Snippet: (A) and (B): RT-qPCR analysis of reactive mRNA of IL-1β and TNF-α expression in hypoxic SCs and the intervention effect of melittin. (C) Immunofluorescence assays after staining with anti-Fn and anti-IL-1R1 antibodies (scale bars = 50 μm). ****p < 0.0001 versus the control group. Data are presented as mean ± standard deviation. SCs: Schwann cells; RT-qPCR: Quantitative reverse transcription-polymerase chain reaction; IL-1R1: Interleukin-1 receptor type 1; TNF-α: Tumor necrosis factor-alpha; DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: Each PVDF membrane was closed with 5% skim milk (BD, Franklin Lakes, NJ) at room temperature for 60 min and then incubated overnight with the following primary antibodies at 4°C: anti-HIF‐1α antibody (#14179, Cell Signaling, 1:1000), anti-GAPDH antibody (ab245355, Abcam, 1:1000), anti-VEGF antibody (#9698, Cell Signaling, 1:1000), anti-IL-1R1 antibody (#6775, Cell Signaling, 1:1000), anti-ENO1 antibody (#3810S, Cell Signaling, 1:1000), anti-C-JUN antibody (#9165T, Cell Signaling, 1:1000), anti-glial cell line-derived neurotrophic factor (GDNF) antibody (#47808, Cell Signaling, 1:1000), anti-IRAK1 antibody (#4395, Cell Signaling, 1:1000), anti-IKKα antibody (#2682 Cell Signaling, 1:1000), anti-p65 antibody (ab16502, Abcam, 1:800), anti-p50 antibody (ab305263, Abcam, 1:900), anti-IKB-α antibody (#8943, Cell Signaling, 1:900), anti-NGF antibody (#2046, Cell Signaling, 1:1000), and anti-β-actin antibody (ab179467, Abcam, 1:1000).

Techniques: Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Control, Standard Deviation, Reverse Transcription, Polymerase Chain Reaction

Journal: Cureus

Article Title: Melittin Alleviates Oxidative Stress Injury in Schwann Cells by Targeting Interleukin-1 Receptor Type 1 to Downregulate Nuclear Factor Kappa B-Mediated Inflammatory Response In Vitro

doi: 10.7759/cureus.65721

Figure Lengend Snippet: (A) RT-qPCR results showing the expression levels of IL-1R1 and IL-1β. (B) Transmission electron microscope images of ultrastructural alterations in CoCl 2 -induced SCs for each group (Left: ×2500, Right: ×7000), with the red arrows indicating mitochondria. (C) Western blot of C-JUN and GDNF protein expression for quantitative analyses. (D) and (E) Western blot and RT-qPCR results showing the expression levels of VEGF, HIF-1α, IL-1R1, ENO1, AR, SOD, NGF, and iNOS. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 versus control and other groups. Data are presented as mean ± standard deviation. SCs: Schwann cells; GDNF: Glial cell line-derived neurotrophic factor; RT-qPCR: Quantitative reverse transcription-polymerase chain reaction; VEGF: Vascular endothelial growth factor; HIF-1α: Hypoxia-inducible factor 1-alpha; IL-1R1: Interleukin-1 receptor type 1; ENO1: Enolase 1; AR: Aldose reductase; SOD: Superoxide dismutase; NGF: Nerve growth factor; iNOS: Inducible nitric oxide synthase.

Article Snippet: Each PVDF membrane was closed with 5% skim milk (BD, Franklin Lakes, NJ) at room temperature for 60 min and then incubated overnight with the following primary antibodies at 4°C: anti-HIF‐1α antibody (#14179, Cell Signaling, 1:1000), anti-GAPDH antibody (ab245355, Abcam, 1:1000), anti-VEGF antibody (#9698, Cell Signaling, 1:1000), anti-IL-1R1 antibody (#6775, Cell Signaling, 1:1000), anti-ENO1 antibody (#3810S, Cell Signaling, 1:1000), anti-C-JUN antibody (#9165T, Cell Signaling, 1:1000), anti-glial cell line-derived neurotrophic factor (GDNF) antibody (#47808, Cell Signaling, 1:1000), anti-IRAK1 antibody (#4395, Cell Signaling, 1:1000), anti-IKKα antibody (#2682 Cell Signaling, 1:1000), anti-p65 antibody (ab16502, Abcam, 1:800), anti-p50 antibody (ab305263, Abcam, 1:900), anti-IKB-α antibody (#8943, Cell Signaling, 1:900), anti-NGF antibody (#2046, Cell Signaling, 1:1000), and anti-β-actin antibody (ab179467, Abcam, 1:1000).

Techniques: Quantitative RT-PCR, Expressing, Transmission Assay, Microscopy, Western Blot, Control, Standard Deviation, Derivative Assay, Reverse Transcription, Polymerase Chain Reaction

Journal: Cureus

Article Title: Melittin Alleviates Oxidative Stress Injury in Schwann Cells by Targeting Interleukin-1 Receptor Type 1 to Downregulate Nuclear Factor Kappa B-Mediated Inflammatory Response In Vitro

doi: 10.7759/cureus.65721

Figure Lengend Snippet: (A-E) RT-qPCR results showing mRNA expressions of IKK, IKB-α, p65, p60, and IRAK1. (F) Western blot results of protein expression of IKK, IKB-α, p65, p60, and IRAK1. (G-K) Quantitative analyses of IKK, IKB-α, p65, p60, and IRAK1 protein expression. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 versus control and other groups. Data are presented as mean ± standard deviation. RT-qPCR: Quantitative reverse transcription-polymerase chain reaction; IL-1R1: Interleukin-1 receptor type 1; NF-κB: Nuclear factor kappa B.

Article Snippet: Each PVDF membrane was closed with 5% skim milk (BD, Franklin Lakes, NJ) at room temperature for 60 min and then incubated overnight with the following primary antibodies at 4°C: anti-HIF‐1α antibody (#14179, Cell Signaling, 1:1000), anti-GAPDH antibody (ab245355, Abcam, 1:1000), anti-VEGF antibody (#9698, Cell Signaling, 1:1000), anti-IL-1R1 antibody (#6775, Cell Signaling, 1:1000), anti-ENO1 antibody (#3810S, Cell Signaling, 1:1000), anti-C-JUN antibody (#9165T, Cell Signaling, 1:1000), anti-glial cell line-derived neurotrophic factor (GDNF) antibody (#47808, Cell Signaling, 1:1000), anti-IRAK1 antibody (#4395, Cell Signaling, 1:1000), anti-IKKα antibody (#2682 Cell Signaling, 1:1000), anti-p65 antibody (ab16502, Abcam, 1:800), anti-p50 antibody (ab305263, Abcam, 1:900), anti-IKB-α antibody (#8943, Cell Signaling, 1:900), anti-NGF antibody (#2046, Cell Signaling, 1:1000), and anti-β-actin antibody (ab179467, Abcam, 1:1000).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Control, Standard Deviation, Reverse Transcription, Polymerase Chain Reaction