monoclonal mouse antibody against il 35p35  (R&D Systems)

Journal: Journal of Virology

Article Title: Hepatitis B Virus Core Particles Containing a Conserved Region of the G Protein Combined with Interleukin-35 Protected Mice against Respiratory Syncytial Virus Infection without Vaccine-Enhanced Immunopathology

doi: 10.1128/JVI.00007-20

Figure Lengend Snippet: Expression of constructed IL-35 in 293T cells and the mRNA levels of Il6 and Il8 in MLFs, MLF/phage, and MLF/phage-IL-35 stimulated with VSV. (A) The sequence of Ebi3 and p35, linked by 3×GGGGS, was inserted into the NheI/KpnI sites of pVAX to generate pIL-35. (B) Western blots were performed to measure IL-35 levels in 293T cells that had been transfected with pVAX or pIL-35 for 48 h. IL-35 was detected with anti-mouse EBI3 or anti-mouse p35 antibody (the molecular mass of IL-35 is ∼50 kDa). (C) Ebi3 and p35 mRNA levels in MLFs, MLF/phage, and MLF/phage-IL-35. The mRNA level was determined from the threshold cycle (CT) values of the PCR amplification. Low threshold cycle values indicate high mRNA levels, while high threshold cycle values represent low mRNA levels. (D) IL-35 expression in MLFs was confirmed by Western blotting with anti-mouse EBI3 antibody and anti-mouse p35 antibody (the molecular mass of IL-35 is ∼50 kDa). (E and F) qRT-PCR analysis of Il6 (E) and Il8 (F) mRNA levels in MLFs, MLF/phage, and MLF/phage-IL-35 after VSV stimulation for 6 h and 12 h. Pairwise comparisons of the values from 3 experiments were performed using a t test. ***, P < 0.01; **, P < 0.01; *, P < 0.05; ns, not significant. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Rel., relative.

Article Snippet: Briefly, the expressed protein samples were separated on 12% polyacrylamide gels, which were transferred onto a polyvinylidene difluoride membrane for Western blot analysis using a mouse anti-EBI3 monoclonal antibody (Rockland Inc., Limerick, PA, USA) and a mouse anti-p35 monoclonal antibody (R&D Systems Inc., Minneapolis, MN, USA).

Techniques: Expressing, Construct, Sequencing, Western Blot, Transfection, Amplification, Quantitative RT-PCR

Journal: Journal of Virology

Article Title: Hepatitis B Virus Core Particles Containing a Conserved Region of the G Protein Combined with Interleukin-35 Protected Mice against Respiratory Syncytial Virus Infection without Vaccine-Enhanced Immunopathology

doi: 10.1128/JVI.00007-20

Figure Lengend Snippet: Primers used in the experiment

Article Snippet: Briefly, the expressed protein samples were separated on 12% polyacrylamide gels, which were transferred onto a polyvinylidene difluoride membrane for Western blot analysis using a mouse anti-EBI3 monoclonal antibody (Rockland Inc., Limerick, PA, USA) and a mouse anti-p35 monoclonal antibody (R&D Systems Inc., Minneapolis, MN, USA).

Techniques: Sequencing