rabbit polyclonal anti hbsag antibody  (Novus Biologicals)

Journal: Frontiers in Microbiology

Article Title: Transfection of unmodified oligodeoxynucleotide with polyethylenimine reduces the level of hepatitis B surface antigen

doi: 10.3389/fmicb.2025.1600679

Figure Lengend Snippet: PEI/40 nt-AC complex selectively reduces the secretion of HBsAg in HepAD38 cells. (A) The gel retardation assay indicated that PEI complexing to 40 nt length oligonucleotide (adenosine-cytosine repeated sequences) at different weight ratio as indicated. 1 μL of 40 nt-AC water solution (100 μM) contains approximately 1.2 ug of 40 nt-AC oligonucleotide. (B) Internalization of FITC-labeled oligonucleotides in HepAD38 cells with fluorescent vision (up) and bright field (down). (C) The cytotoxicity of PEI/oligonucleotide was measured by using the Cell Counting Kit-8 assay. HepAD38 cells were cultured in 12-well plate and transfected with PEI/ON. The culture supernatants were collected to detect HBsAg (D) , HBeAg (E) , and HBV DNA (F) levels after 48 h. Means and SDs of data from three independent experiments are plotted. (G) HepAD38 cells were cultured in 6-well plate and transfected with PEI/ON as indicated. Cells were lysed with 100 μL of RIPA lysis buffer (25 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS). The intracellular levels of HBsAg and HBeAg were determined. (H) The replication intermediates in viral core particles were examined by Southern blot and Viral RNAs by Northern blot. RC, relaxed circular DNA. 18S/28S rRNAs served as the RNA loading control. Means and SDs of data from at least three independent tests were plotted. *** p < 0.001.

Article Snippet: Polyclonal rabbit anti-Hepatitis B virus core antigen (HBc) antibody (B0586) was purchased from the Dako (Glostrup, Denmark), polyclonal rabbit anti-HBsAg antibody (NB100-62652) from Novus Biological (Littleton, CO, USA) and β-actin (AF0003) was purchased from the Beyotime Biotechnology (Shanghai, China).

Techniques: Electrophoretic Mobility Shift Assay, Labeling, Cell Counting, Cell Culture, Transfection, Lysis, Southern Blot, Northern Blot, Control

Journal: Frontiers in Microbiology

Article Title: Transfection of unmodified oligodeoxynucleotide with polyethylenimine reduces the level of hepatitis B surface antigen

doi: 10.3389/fmicb.2025.1600679

Figure Lengend Snippet: PEI/pEGFP-C2 complexes have no impact on the expression levels of HBsAg. HepAD38 cells were cultured in 12-well plate and treated with the PEI (A) or ON (B) alone and the culture supernatants were collected to detect HBsAg, HBeAg after 48 h. (C) Green fluorescent protein (GFP) expression in cells in a dose dependent manner with fluorescent vision (up) and bright field (down) by fluorescence microscope. (D) The gel retardation assay indicated that PEI complexing to pEGFP-C2 at different weight ratio as indicated. The culture supernatants were collected to detect HBsAg, HBeAg (E) and HBV DNA level (F) after 48 h of transfection. Means and SDs of data from three independent experiments are plotted.

Article Snippet: Polyclonal rabbit anti-Hepatitis B virus core antigen (HBc) antibody (B0586) was purchased from the Dako (Glostrup, Denmark), polyclonal rabbit anti-HBsAg antibody (NB100-62652) from Novus Biological (Littleton, CO, USA) and β-actin (AF0003) was purchased from the Beyotime Biotechnology (Shanghai, China).

Techniques: Expressing, Cell Culture, Fluorescence, Microscopy, Electrophoretic Mobility Shift Assay, Transfection

Journal: Frontiers in Microbiology

Article Title: Transfection of unmodified oligodeoxynucleotide with polyethylenimine reduces the level of hepatitis B surface antigen

doi: 10.3389/fmicb.2025.1600679

Figure Lengend Snippet: PEI/ON complexes inhibit secreted HBsAg in a size dependent manner. HepAD38 cells cultured in 12-well plate were transfected with unmodified oligodeoxynucleotides (100 μM) in different indicated nt-length using PEI (1 μg/μL). (A) The oligodeoxynucleotide in 10 nt length, 10 nt-AC. (B) The oligodeoxynucleotide in 20 nt length, 20 nt-AC. (C) The oligodeoxynucleotide in 30 nt length, 30 nt-AC. (D) The oligodeoxynucleotide in 40 nt length, 40 nt-AC. Post 48 h transfection, the culture supernatants were collected to detect HBsAg, HBeAg. Means and SDs of data from three independent experiments are plotted. * p < 0.05, ** p < 0.01.

Article Snippet: Polyclonal rabbit anti-Hepatitis B virus core antigen (HBc) antibody (B0586) was purchased from the Dako (Glostrup, Denmark), polyclonal rabbit anti-HBsAg antibody (NB100-62652) from Novus Biological (Littleton, CO, USA) and β-actin (AF0003) was purchased from the Beyotime Biotechnology (Shanghai, China).

Techniques: Cell Culture, Transfection

Journal: Frontiers in Microbiology

Article Title: Transfection of unmodified oligodeoxynucleotide with polyethylenimine reduces the level of hepatitis B surface antigen

doi: 10.3389/fmicb.2025.1600679

Figure Lengend Snippet: PEI/ON complexes uniquely inhibit HBsAg secretion comparing with other commercial transfection reagents. HepAD38 cells cultured in 12-well plate were transfected with 40 nt-AC oligonucleotides using the indicated commercial transfection reagents. 48 h post of transfection, internalization of FITC-labeled oligonucleotides in HepAD38 cells in fluorescent vision (up) and white field (down) by using different commercial transfection reagents. (A) PEI. (B) OneStep transfecter: One-step DNA Transfecter (ENLIGHTEN Biotech, LD210-005). (C) X-tremeGENE: X-tremeGENE 9 DNA Transfection Reagent (Roche, 06365779001). (D–F) The culture supernatants were collected to detect HBsAg and HBeAg levels after 48 h of transfection. **** p < 0.0001.

Article Snippet: Polyclonal rabbit anti-Hepatitis B virus core antigen (HBc) antibody (B0586) was purchased from the Dako (Glostrup, Denmark), polyclonal rabbit anti-HBsAg antibody (NB100-62652) from Novus Biological (Littleton, CO, USA) and β-actin (AF0003) was purchased from the Beyotime Biotechnology (Shanghai, China).

Techniques: Transfection, Cell Culture, Labeling

Journal: Frontiers in Microbiology

Article Title: Transfection of unmodified oligodeoxynucleotide with polyethylenimine reduces the level of hepatitis B surface antigen

doi: 10.3389/fmicb.2025.1600679

Figure Lengend Snippet: PEI/Decoy-ON inhibits HBsAg secretion levels but not HBV transcripts. HepAD38 cells cultured in 12-well plate were co-transfected with scramble oligonucleotides with different initial weight ratios (PEI: ON) as indicated. The culture supernatants were collected to detect HBsAg and HBeAg levels (A) and HBV DNA levels (B) after 48 h of transfection. (C) HepAD38 cells were cultured in 6-well plate and transfected with scramble ON using PEI as indicated. HBV replication intermediates in viral core particles were examined with Southern blot and Viral RNAs with Northern blot. (D) The HBsAg levels (up) and HBeAg levels (down) in supernatants of cells transfected with the scramble double-stranded oligonucleotide or decoy oligonucleotides of Sp1, CREB and HNF1α, respectively. Sp1, Specificity protein 1; HNF1α, hepatocyte nuclear factor 1α; CREB, C-AMP-response element binding protein. (E) HepAD38 cells cultured in 6-well plate were transfected with PEI/decoy-ON as indicated. HBV replication intermediates in viral core particles were examined with Southern blot and Viral RNAs with Northern blot. rcDNA, relaxed circular DNA. 18S/28S RNAs served as the RNA loading control. Means and SDs of data from three independent experiments are plotted. ** p < 0.01, *** p < 0.001.

Article Snippet: Polyclonal rabbit anti-Hepatitis B virus core antigen (HBc) antibody (B0586) was purchased from the Dako (Glostrup, Denmark), polyclonal rabbit anti-HBsAg antibody (NB100-62652) from Novus Biological (Littleton, CO, USA) and β-actin (AF0003) was purchased from the Beyotime Biotechnology (Shanghai, China).

Techniques: Cell Culture, Transfection, Southern Blot, Northern Blot, Binding Assay, Control

Journal: Frontiers in Microbiology

Article Title: Transfection of unmodified oligodeoxynucleotide with polyethylenimine reduces the level of hepatitis B surface antigen

doi: 10.3389/fmicb.2025.1600679

Figure Lengend Snippet: PEI/oligonucleotide transfection prior to viral inoculation inhibits HBV infection in HepG2-NTCP cells. (A) Treatment procedure: HepG2-NTCP cells were transfected by the indicated ratios of PEI/ONs for 5 h, following by 12 h of inoculation with HBV at 1,000 GE per cell. Supernatants were harvested for secreted HBsAg, HBeAg in day 0, 4, and 8 post-inoculation. (B) The culture supernatants were collected to detect both secreted HBsAg and HBeAg at day 0, 4 and 8 post-infection. Dotted lines represent cut-off thresholds. (C) Post 8 day infection, HBcAg were detected by anti-HBc with fluorescent tag (red) in cells in which nuclei were labeled with DAPI (blue) by fluorescence microscopy. The exact numbers of DPAI dying and HBcAg (+) cells were analyzed and summarized by Image J software. All data, expressed as means ±standard deviation, were independently reproduced two times.

Article Snippet: Polyclonal rabbit anti-Hepatitis B virus core antigen (HBc) antibody (B0586) was purchased from the Dako (Glostrup, Denmark), polyclonal rabbit anti-HBsAg antibody (NB100-62652) from Novus Biological (Littleton, CO, USA) and β-actin (AF0003) was purchased from the Beyotime Biotechnology (Shanghai, China).

Techniques: Transfection, Infection, Labeling, Fluorescence, Microscopy, Software, Standard Deviation

Journal: Frontiers in Microbiology

Article Title: Transfection of unmodified oligodeoxynucleotide with polyethylenimine reduces the level of hepatitis B surface antigen

doi: 10.3389/fmicb.2025.1600679

Figure Lengend Snippet: PEI/40 nt-AC complex reduces HBsAg level in the HDI BALB/c mice. (A) Schematic representation of the mice experiment. The 6-weeks-old BALB/c male mice ( n = 6/group) were hydrodynamiclly injected with pHBV1.3 plasmid through tail-vein at day 0. The injections of PBS, PEI, PEI/10 nt-AC complex and PEI/40 nt-AC complex are indicated by red arrows. (B) Serum samples were collected at indicated time points and the levels of serum HBsAg and HBeAg kinetics in mice were analyzed. Cut-off thresholds of determination are 0.05 IU/mL for HBsAg and 1.000 S/CO for HBeAg, which are represented by dotted lines. The positive rates of serum HBsAg and HBeAg in mice were analyzed and presented (right panel). (C) Sera were collected at 42 days post of pHBV1.3-injection and the levels of anti-HBsAg (left) and anti-HBeAg (right) were analyzed. (D) Serum levels of alanine transaminase (ALT) and aspartate transaminase (AST) were measured. A two-tailed Student t test was used. * p < 0.05. (E) Mice were sacrificed at 42 days post of pHBV1.3 injection. Liver sections taken from HDI-mice in different groups were stained for HBsAg (black arrow heads) in addition to (F) HE staining. Representative images from control and PEI/ON transfected mice are shown. Scale bar = 250 μm.

Article Snippet: Polyclonal rabbit anti-Hepatitis B virus core antigen (HBc) antibody (B0586) was purchased from the Dako (Glostrup, Denmark), polyclonal rabbit anti-HBsAg antibody (NB100-62652) from Novus Biological (Littleton, CO, USA) and β-actin (AF0003) was purchased from the Beyotime Biotechnology (Shanghai, China).

Techniques: Injection, Plasmid Preparation, Two Tailed Test, Staining, Control, Transfection