human ifn λ3 duoset elisa kits  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: Next-generation single-cycle respiratory syncytial virus vaccines with increased type I interferon induction yield robust systemic and mucosal responses in mice

doi: 10.3389/fimmu.2026.1778423

Figure Lengend Snippet: Impact of reduced NS1 levels on IFN expression. A549 cells were infected with RSV-Mnull/G1, RSV-Mnull/F1, RSV-Mnull, and RSV-rWT, or left uninfected (mock). Infected cell supernatant was examined at 24 hpi. (A) mRNA levels. Total RNA was extracted from infected cells and real-time PCR was performed (primers listed in Materials and Methods). IFN levels were normalized to RSV N protein mRNA levels. Fold change is indicated on the Y-axis. (B) Cytokine ELISA. Supernatants were harvested from infected cells, and cytokine levels were determined using commercially available human IFN-β or IFN-λ3 specific kits. Error bars are the standard deviation of the mean of triplicate samples. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: The levels of IFN-β and IFN-λ3 in the supernatants were determined using the human IFN-β and human IFN-λ3 DuoSet ELISA kits (R&D Systems, MN, USA), following manufacturer’s instructions.

Techniques: Expressing, Infection, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: bioRxiv

Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling

doi: 10.64898/2026.03.24.712780

Figure Lengend Snippet: ( A ) HEK293T reporter cells were treated with 6.0 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle, cells were imaged by confocal microscopy at 6 h post treatment to examine conjugate colocalization with target TBK1 (representative of N = 3 biological replicates). Scale bar is 10 μm. ( B ) HEK293T reporter cells were treated with 8.3 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle, Western blot was performed at 6 h post treatment to examine TBK1 and IRF3 phosphorylation (representative of N = 3 biological replicates). ( C ) IRF3 reporter signal relative to buffer treatment for HEK293T reporter cells pretreated for 6 h with TBK1 inhibitor MRT67307 (TBK1i) and then treated with 8.3 μg/mL STING or Scr conjugate delivered using TransIT-X2, measured 24 h post treatment (N = 3 biological replicates). ( D ) Western blot of STING and β-actin expression in ovarian cancer cell lines KURAMOCHI and A2780. ( E-F ) KURAMOCHI and A2780 ovarian cancer cell lines were treated with 5.0 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle or 50 μM STING agonist ADU-S100. ( E ) CXCL10 and ( F ) IFN-β in supernatant was measured by ELISA 24 h post treatment (N = 3 biological replicates). Replicates where analyte was below the limit of detection (LOD) are labeled as not detected (ND), no summary statistics were computed if any replicate was ND. ( G-I ) KURAMOCHI cells were treated with 5.0 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle or 50 μM ADU-S100, mRNA sequencing was performed at 6 h post treatment (N = 4 biological replicates). ( G ) Plot of Log2 fold change of STING conjugate or ADU-S100 treatment compared to Buffer, showing high correlation between treatments. Plot of Log2 fold change of Scr conjugate or ADU-S100 treatment compared to Buffer is displayed below as a control, showing greatly reduced correlation. The coefficient of determination R 2 for line of best fit is displayed. ( H ) Gene set enrichment analysis was performed on MSigDB Hallmark gene set, normalized enrichment and adjusted P value are displayed for the 10 gene sets significantly enriched ( P < .05) when comparing STING to Scr conjugate. Normalized enrichment and adjusted P values for the same 10 gene sets are displayed for the comparison of ADU-S100 to Buffer ( I ) Heatmap of gene expression for selected genes. Replicates where a given gene was not detected are labeled ND. Data represented as geometric mean ± SD.

Article Snippet: DuoSet ELISA kits (R&D Systems) for human CXCL10 (#DY266), human IFN-β (#DY814), mouse CXCL10 (#DY466), mouse IFN-β (#DY8234), mouse IL-6 (#DY406), mouse TNF-α (#DY410), and mouse IFN-γ (#DY485) were used with 1-Step TMB ELISA Substrate Solution (Thermo Scientific) following vendor instructions.

Techniques: Confocal Microscopy, Western Blot, Phospho-proteomics, Expressing, Enzyme-linked Immunosorbent Assay, Labeling, Sequencing, Control, Comparison, Gene Expression

Journal: bioRxiv

Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling

doi: 10.64898/2026.03.24.712780

Figure Lengend Snippet: ( A-B ) Mice were dosed with 20 μg of STING conjugate delivered by LNP IP and serum was collected at 0, 1, 3, 6, 10, 24, and 50 h. Serum was analyzed to measure ( A ) STING conjugate concentration by Cy5 fluorescence (showing one phase exponential decay fit to data) and ( B ) CXCL10 concentration by ELISA (N = 3 mice). Conditions where analyte was below the limit of detection (LOD) are labeled as not detected (ND). ( C ) Mice were inoculated with 3×10 6 BPPNM cells IP and dosed with 20 μg of STING or Scr conjugate (N = 3 mice) delivered by LNP IP at 14 days after inoculation. ( D ) Omental tumor, ( E ) ascites, and ( F ) serum were collected 6 h after dosing. Concentrations of CXCL10, IFN-β, IL-6, TNF-α, and IFN-γ were measured by ELISA and are reported relative to total protein concentration in tumor and ascites ( D-E ) or relative to volume in serum ( F ). Conditions where analyte was below the LOD are labeled as ND. Average fold change increases in cytokine concentration for STING conjugate treatment compared to Scr conjugate are displayed. Where cytokine level was ND, a lower bound on the fold change was computed by setting all ND replicates as the LOD. Data represented as mean ± SD.

Article Snippet: DuoSet ELISA kits (R&D Systems) for human CXCL10 (#DY266), human IFN-β (#DY814), mouse CXCL10 (#DY466), mouse IFN-β (#DY8234), mouse IL-6 (#DY406), mouse TNF-α (#DY410), and mouse IFN-γ (#DY485) were used with 1-Step TMB ELISA Substrate Solution (Thermo Scientific) following vendor instructions.

Techniques: Concentration Assay, Fluorescence, Enzyme-linked Immunosorbent Assay, Labeling, Protein Concentration

Journal: bioRxiv

Article Title: The human DEAD-box protein DDX3X regulates host and viral mRNA translation during Sendai Virus infection

doi: 10.64898/2026.03.08.707086

Figure Lengend Snippet: (A) Levels of IFNB secreted by NSC and shDDX3X HEK293T cells either uninfected or infected for 16h with SeV, with 72h DDX3X knockdown, measured via ELISA (n = 6). DDX3X knockdown and even levels of β-actin between NSC and shDDX3X samples were confirmed via western blot of cell lysates as shown in (C). (B) Western blot of HEK293T cells infected with SeV for varying lengths of time, probed with anti-SeV polyclonal antibody. Approximate apparent molecular weights, as well as predicted protein identities of bands, are shown. (C) Western blot using anti-SeV pAb, DDX3X and β-actin antibody on samples from DDX3X knockdown and control HEK293Ts infected for various times with SeV. The sample shown in the representative blot corresponds to one of the biological replicates in (A). 2-3 biological replicates (dependent on time point) were carried out and quantified as shown in Figure S3.

Article Snippet: For ELISAs, undiluted cell culture supernatants collected from NSC and shDDX3X HEK293T cells after 72 h knockdown and (where indicated) 16hpi with SeV were assayed using a Human IFN-beta ELISA kit (R&D systems, 41410-1), following manufacturer’s instructions.

Techniques: Infection, Knockdown, Enzyme-linked Immunosorbent Assay, Western Blot, Control