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: SARS-CoV-2 Defective Viral Genomes from Distinct Genomic Regions Drive Divergent Interferon Responses

doi: 10.64898/2026.03.19.712870

Figure Lengend Snippet: (A-B) A549-ACE2 cells were infected with SARS-CoV-2 (MOI 0.8), DVG-B ( A , MOI 1), DVG-A ( B , MOI 1) or co-infected with either DVG-B:SARS-CoV-2 (MOI 1: MOI 0.8) or DVG-A: SARS-CoV-2 (MOI 1: MOI 0.8) for 24 hours post-inoculation. Expression of host genes and viral genes/specific DVGs were calculated relative to GAPDH by RT-qPCR. *p<0.05, **p<0.01 by two-way ANOVA with Holm-Sidak’s multiple comparisons test. N=4, mean±SD ( C ) Supernatants from mock, DVG-B, SARS-CoV-2 and DVG-B/SARS-CoV-2 co-infected cells were collected and secreted IFNB1 was measured by ELISA. N=3, mean±SD. ( D ) Progeny infectious virus production was determined from supernatants of SARS-CoV-2, DVG-B/SARS-CoV-2 and DVG-A/SARS-CoV-2 co-infected cells (A-B) by TCID 50 /mL. Viral titration was performed in VeroAT cells. *p<0.05 by paired one-tailed t-test. N=4, mean±SD. ( E-F ) Cells were infected with SARS-CoV-2 (MOI 0.1) with increasing ratios of DVG-B: SARS-CoV-2 ( E , MOI 0.1, 0.5, 2.5: MOI 0.1) or DVG-A: SARS-CoV-2 ( F , MOI 0.1, 0.2, 0.5, 1: MOI 0.1). IFNB1, N and specific DVG copy numbers were quantified relative to GAPDH by RT-qPCR. SARS-CoV-2 titers, with increasing input MOIs of DVG-B or DVG-A, were measured by TCID 50 /mL. *p<0.05, **p<0.01, ***p<0.001 by one-way ANOVA with Dunnett’s multiple comparisons test. N=2, mean±SD.

Article Snippet: IFNB1 concentrations within the supernatants of SARS-CoV-2, DVG-B and SARS-CoV-2/DVG-B co-infected cells were determined using the VeriKine-HS Human Interferon-Beta TCM ELISA Kit (PBL Assay Science catalog no. 41435) according to the manufacturer’s instructions.

Techniques: Infection, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Virus, Titration, One-tailed Test