elisa kit #e-el-m1084c  (Elabscience Biotechnology)

Journal: eLife

Article Title: Vangl2 suppresses NF-κB signaling and ameliorates sepsis by targeting p65 for NDP52-mediated autophagic degradation

doi: 10.7554/eLife.87935

Figure Lengend Snippet: ( A ) Transcription levels of Vangl2 in PBMCs from healthy volunteers (healthy control, HC) and sepsis patients were analyzed by real-time PCR ( n = 8). ( B ) Vangl2 mRNA in different organs from mice treated with or without LPS ( n ≥ 3). ( C ) The survival rates of wild-type (WT) and Vangl2 ΔM mice treated with high dosage of LPS (30 mg/kg, intraperitoneally [i.p.]) ( n ≥ 3). ( D–F ) WT and Vangl2 ΔM mice ( n ≥ 3) were treated with LPS (30 mg/kg, i.p.). Splenocytes were collected at 9 hr after LPS treatment. Cell lysates of CD11b + sorted splenocytes were analyzed by immunoblotting with the indicated antibodies ( n ≥ 3) ( D ). RNAs from splenocytes were isolated and used for expression analysis of Il1b , Tnfa , and Il6 using qPCR ( n ≥ 3) ( E ). Sera were collected at indicated times post LPS treatment and subjected to enzyme-linked immunosorbent assay (ELISA) analysis of IL-1β, tumor necrosis factor-α (TNF-α), and IL-6 ( n ≥ 3) ( F ). PBMCs, peripheral blood mononuclear cells; Unsti, unstimulation; LPS, lipopolysaccharide; LN, lymph node; SP, spleen. Data are representative of three independent experiments and are plotted as the mean ± standard deviation (SD). Two-tailed Student's t test for A. Multiple t tests for B, E and F. Log-rank (Mantel-Cox) test for survival curve. *p < 0.05, **p < 0.01, ***p < 0.001 vs. corresponding control. Figure 1—source data 1. Uncropped and labelled gels for . Figure 1—source data 2. Raw data for . Figure 1—source data 3. Raw unedited gels for .

Article Snippet: IL-1β, IL-6, and TNF-α in cell supernatants and mice serum were measured using ELISA kit (#E-EL-M0037c, #E-EL-M0044c, and #E-EL-M1084c, respectively; Elabscience Biotechnology) following the manufacturer’s instructions.

Techniques: Control, Real-time Polymerase Chain Reaction, Western Blot, Isolation, Expressing, Enzyme-linked Immunosorbent Assay, Standard Deviation, Two Tailed Test

Journal: eLife

Article Title: Vangl2 suppresses NF-κB signaling and ameliorates sepsis by targeting p65 for NDP52-mediated autophagic degradation

doi: 10.7554/eLife.87935

Figure Lengend Snippet: Wild-type (WT) and Vangl2-deficient ( n ≥ 3) pMAC ( A, C ) or neutrophils ( B, D ) were stimulated with LPS (100 ng/ml) for the indicated times. Immunoblot analysis of total and phosphorylated p65, IKKα/β ( A, B ), and analysis of gray intensity was shown ( C, D ) ( n ≥ 3). ( E, F ) WT and Vangl2-deficient ( n ≥ 3) pMAC or neutrophils were stimulated with LPS (100 ng/ml) for 6 hr. mRNA levels of Il6 and Tnfa were measure by qPCR ( E ). IL-6 and tumor necrosis factor-α (TNF-α) secretion by WT and Vangl2-deficient bone marrow-derived macrophages (BMDMs) or neutrophils treated with or without LPS for 6 hr was measured by enzyme-linked immunosorbent assay (ELISA) ( F ) ( n ≥ 3). ( G, H ) The WT and Vangl2-deficient ( n ≥ 3) neutrophils were treated with LPS (1000 ng/ml) for 4 hr, and the nuclear translocation of p65 was detected by immunofluorescence ( G ) (p65, green; 4'-6-diamidino-2-phenylindole (DAPI), blue; scale bar, 50 μm). Percentages of p65 nuclear translocated cells in WT and Vangl2-deficient neutrophils were determined by counting 100–150 cells in non-overlapping fields ( H ) ( n ≥ 3). ( I, J ) A549 cells were transfected with Flag-tagged Vangl2 plasmid or empty vector, then stimulated with LPS (100 ng/ml) for the indicated times. Immunoblot analysis of total and phosphorylated p65, IKKα/β ( I ) (representative image), and analysis of gray intensity was shown ( J ) ( n ≥ 3). pMAC, peritoneal macrophage; Neu, neutrophil; EV, empty vector. Data are representative of three independent experiments and are plotted as the mean ± standard deviation (SD). Multiple t tests for C, D, E, F and J. Two-tailed Student's t test for H. *p < 0.05, ***p < 0.001 vs. corresponding control. Figure 2—source data 1. Uncropped and labelled gels for . Figure 2—source data 2. Raw data for . Figure 2—source data 3. Raw unedited gels for .

Article Snippet: IL-1β, IL-6, and TNF-α in cell supernatants and mice serum were measured using ELISA kit (#E-EL-M0037c, #E-EL-M0044c, and #E-EL-M1084c, respectively; Elabscience Biotechnology) following the manufacturer’s instructions.

Techniques: Western Blot, Derivative Assay, Enzyme-linked Immunosorbent Assay, Translocation Assay, Immunofluorescence, Transfection, Plasmid Preparation, Standard Deviation, Two Tailed Test, Control

Journal: Nutrients

Article Title: Friedelin Alleviates the Pathogenesis of Collagenase-Induced Tendinopathy in Mice by Promoting the Selective Autophagic Degradation of p65

doi: 10.3390/nu14081673

Figure Lengend Snippet: The resource of Reagents and Antibodies.

Article Snippet: Anti-TNF-α (ELISA) , Elabscience Biotechnology , #E-EL-M1084c.

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, CCK-8 Assay