human hbsag elisa kit  (Novus Biologicals)

Journal: iScience

Article Title: A key role for the exoribonuclease XRN1 in regulating the hepatitis B viral transcriptome

doi: 10.1016/j.isci.2026.116328

Figure Lengend Snippet: XRN1 limits HBV RNAs (A) HBV infection and pAp treatment. Arrested HepG2-NTCP cells were infected with HBV for 16 h, inoculum removed, and cells treated with vehicle or 4 mM 3′, 5′-bisphosphate (pAp) for 72 h. RNA was extracted and total HBV RNAs quantified by RT-qPCR, normalized to the housekeeping gene B2M and expressed relative to vehicle-treated cells (mean ± SD, n = 9 from 3 independent experiments, one-way ANOVA, with multiple comparisons, ∗∗∗ p < 0.001). Cytotoxicity was evaluated by LDH assay with data expressed relative to cells treated with 0.05% Triton X-100 (Pos). (B) HBV infection of WT and XRN1 KO cells. WT and KO HepG2-NTCP cells were assessed for XRN1 expression by western blotting and treated with 50 nM Alexa 647 conjugated preS1 2-48 peptide in the presence or absence of excess unlabeled peptide (200 nM). After washing, the cells were incubated with DAPI and visualized by microscopy with a 20 × objective, scale bars, 150 μm. WT and XRN1 KO HepG2-NTCP cells were inoculated with HBV for 16 h. Intracellular DNA was extracted after 16 h post-infection and RNA at 72 h post-infection. HBV DNA, cccDNA, and total HBV RNAs were quantified by qPCR and normalized to the housekeeping genes, PrP or B2M , respectively. Data are plotted relative to the WT cells (mean ± SD, n = 6 from 2 independent experiments). (C) HBV life cycle and kinetic analysis of infection. WT and XRN1 KO HepG2-NTCP cells were inoculated with HBV for 16 h in the presence or absence of preS1 2-48 peptide, unbound virus removed by washing, and the cells cultured for 3, 6, 9, or 10 days. The schematic depicts steps in the HBV life cycle that were evaluated. Intracellular HBV RNA and secreted HBeAg were quantified at 3, 6, and 9 dpi by RT-qPCR and ELISA, respectively. RT-qPCR data are expressed relative to the housekeeping gene, B2M (mean ± SD, n = 9 from 3 independent experiments). Intracellular rcDNA and core-associated HBV DNA were quantified by qPCR (mean ± SD, n = 6 from 3 independent experiments). XRN1, HBs, HBc, and β-Actin proteins were detected at 10 dpi. Statistical significance was determined using Mann-Whitney tests, with Bonferroni correction for multiple comparisons; ∗∗ p < 0.01, ∗∗∗ p < 0.005, and ∗∗∗∗ p < 0.0001. Please see also .

Article Snippet: HBsAg ELISA , Autobio , Cat# CL0310-2.

Techniques: Infection, Quantitative RT-PCR, Lactate Dehydrogenase Assay, Expressing, Western Blot, Incubation, Microscopy, Virus, Cell Culture, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: iScience

Article Title: A key role for the exoribonuclease XRN1 in regulating the hepatitis B viral transcriptome

doi: 10.1016/j.isci.2026.116328

Figure Lengend Snippet: HBV induces P-bodies (A and B) Effect of HBV infection on XRN1 and EDC4 expression. Mock or HBV-infected HepG2-NTCP cells were cultured for up to 9 days (A) P-body components XRN1 and EDC4, together with HBsAg and β-actin, were detected at 3, 6, and 9 dpi by western blotting. (B) EDC4 (green) and DAPI nuclei (blue) were detected by immunofluorescence at 6 dpi, with a representative image shown (scale bars, 25 μm). The average number of EDC4 puncta relative to nuclei was quantified in 9 images from 3 independent experiments, with significance assessed using a Mann-Whitney test (∗∗∗ p < 0.001, ∗ p < 0.05). (C) Abundance of viral and host RNAs. Total HBV RNAs (HBV-total, yellow) and host Peptidylprolyl isomerase B RNA (PPIB, red) and nuclear DAPI (blue) were imaged at 6 dpi (scale bars, 20 μm). (D) Replication of in vitro transcribed HBV pgRNA. A schematic of in vitro transcribed (IVT) WT or Δε pgRNA and their viral products. Mock or IVT HBV RNA-transfected (100 ng) Huh 7.5 cells were cultured for 2, 4, or 6 days. Cells were imaged for HBc (magenta) (scale bars, 75 μm), intracellular HBV DNA, and secreted HBsAg assessed by qPCR or ELISA, respectively. (E) pCpg RNA levels after 6 days were measured by RT-qPCR, and the data were normalized to a housekeeping gene, B2M , respectively. (F) EDC4 puncta in HBV RNA-transfected cells. Mock or IVT HBV RNA-transfected (100 ng) Huh 7.5 cells (6 days) were stained for EDC4 expression and puncta number and size quantified in 15 images from 3 independent experiments. Mean EDC4 values are expressed relative to the number of nuclei per image, with statistical significance assessed using a Mann-Whitney test with correction for multiple comparisons (∗∗∗ p < 0.001 and ∗ p < 0.05).

Article Snippet: HBsAg ELISA , Autobio , Cat# CL0310-2.

Techniques: Infection, Expressing, Cell Culture, Western Blot, Immunofluorescence, MANN-WHITNEY, In Vitro, Transfection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Staining

Journal: Frontiers in Pharmacology

Article Title: Sophora tonkinensis enhances activation of cGAS-STING pathway and restrains HBV replication

doi: 10.3389/fphar.2025.1630460

Figure Lengend Snippet: STE promotes cGAS-STING pathway activation via facilliating 2′3′-cGAMP synthesis (A,B) BMDMs or THP-1 were first treated with STE for 1 h and then with ISD for 2.5 h. Subsequently, cell membranes and nuclei were isolated and the amount of IRF3 protein in them was detected. (C–F) The cGAS, STING, TBK1 or IRF3-5D plasmids were transfected into HEK-293 cells for 18 h, then STE (1 mg/mL) was administered, and the expression level of IFN-β in them was detected by qPCR after 6 h (n = 3). (G) BMDMs were treated with STE for 1.5 h followed by ISD for 1.5 h and 2′3′-cGAMP levels in the samples were detected using ELISA kits (n = 3). (H–J) THP-1 cells inoculated in 24-well plates were first treated with STE. Transwell chambers inoculated with HepG2.2.2.15 cells were placed in well plates containing THP-1 cells, and after 24 to detect the HBsAg and IFN-β levels (n = 3). Data are expressed as mean ± SD from three biological replicates. Statistical analyses were performed on multiple samples using one-way ANOVA with Dunnett’s post hoc test. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. the control, ns, not significant.

Article Snippet: Human hepatitis B virus e antigen (HBeAg) ELISA Kit (ZC01803, Wantai biopharm), Human hepatitisB surface antigen (HBsAg) ELISA Kit (ZCY0503, Wantai biopharm), Quantitative PCR Diagnostic Kit for HBV-DNA (PCR-Fluorescence Probing in One-Tube) (B001, NaGene Diagnosis), ISD was synthesized as previously described by heating equimolar amounts of sense DNA and anti-DNA oligo acids to 95 °C for 5 min and then cooling to room temperature.

Techniques: Activation Assay, Isolation, Transfection, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Pharmacology

Article Title: Sophora tonkinensis enhances activation of cGAS-STING pathway and restrains HBV replication

doi: 10.3389/fphar.2025.1630460

Figure Lengend Snippet: STE restrains HBV replication and promotes cGAS-STING pathway activation. (A–D) The HBV model was constructed by injecting 20 μg of pAAV/HBV1.2 plasmid into mice, and STE or ETV was given daily, while mice in the blank group were given saline (A) . The serum levels of HBsAg (B) , HBeAg (C) and HBV DNA (D) in mice were detected weekly. (E,F) Immunohistochemical method to detect the expression level of HBcAg in fractional liver tissues, scale bars: 250 μm (top row); 50 μm (bottom row) (n = 6). (G,H) Serum levels of ALT and AST were measured by ELISA kits and physiological and biochemical kits (n = 6). Data are presented as mean ± SD (n = 6 per group) one-way ANOVA and Dunnett’s post hoc test were used to assess the differences of multiple groups, ***p < 0.001 vs. the model group, ns, not significant.

Article Snippet: Human hepatitis B virus e antigen (HBeAg) ELISA Kit (ZC01803, Wantai biopharm), Human hepatitisB surface antigen (HBsAg) ELISA Kit (ZCY0503, Wantai biopharm), Quantitative PCR Diagnostic Kit for HBV-DNA (PCR-Fluorescence Probing in One-Tube) (B001, NaGene Diagnosis), ISD was synthesized as previously described by heating equimolar amounts of sense DNA and anti-DNA oligo acids to 95 °C for 5 min and then cooling to room temperature.

Techniques: Activation Assay, Construct, Plasmid Preparation, Saline, Immunohistochemical staining, Expressing, Enzyme-linked Immunosorbent Assay