standard of mouse reference serum  (Bethyl)

Journal: The World Allergy Organization Journal

Article Title: High protein diet increases the risk of allergic sensitization but not asthma in mice through modulation of the cytokine milieu toward Th2 bias ☆

doi: 10.1016/j.waojou.2025.101031

Figure Lengend Snippet: Effect of feeding mice for 7 weeks either normal diet or high protein diet on serum levels of (A) OVA-specific IgE (B) OVA-specific IgG1 and (C) OVA-specific IgG2a from OVA-sensitized mice (n = 11 in each group) after OVA intranasal challenge. The high protein diet group showed higher levels of both OVA-specific IgE and IgG1compared to the normal diet group. ns: not significant, CI: confidence interval. IQR: interquartile range.

Article Snippet: After washing the wells, blocking buffer (0.5% gelatin in wash buffer or 3% Rat serum in wash buffer) was added and the plates incubated for 2 h. Appropriate reference standard OVA-IgE (Chondrex #3006) or OVA-IgG1 (Chondrex # 7093) or OVA-IgG2a (Chondrex # 7095) and diluted serum samples were then added to the wells and incubated for 3 h. This was followed by incubation with OVA-Biotin for 1 h and with streptavidin-HRP for 30 min. Tetramethylbenzidine substrate was added and the enzyme-substrate reaction stopped after 15 min, and the optical density measured at 450 nm.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Surface Engineering of HEK293 Cell-Derived Extracellular Vesicles for Improved Pharmacokinetic Profile and Targeted Delivery of IL-12 for the Treatment of Hepatocellular Carcinoma

doi: 10.2147/IJN.S388916

Figure Lengend Snippet: Examination of the effect of mouse single chain IL12 and HN3 nanobody on the cell-binding properties of EVs. ( A ) Schematic design of an exosomal nanobody (HN3) and mouse single chain IL12 (mscIL12), whose vesicular loading was driven by the transmembrane domains of PDGFRβ and ITGB1, respectively. ( B ) Dose-response curve of mscIL12 EVs in mouse splenocytes, analyzed using the “drc” package in R. ( C ) Representative fluorescence images of HepG2 cells after incubation with PKH67-labeled EVs. Nuclei were stained with DAPI. ( D ) Cellular NanoLuc activities in various cell lines after incubation with ITGB1 − mscIL12 + HN3 + EVs, or ITGB1 − mscIL12 + HN3 + EVs in the presence of hGPC3-Fc fusion protein. Luciferase activities were normalized to the chemiluminescence intensities of cells incubated with respective EVs alone. ( E ) Particle populations (normalized to the plasma concentration) of EVs with or without the GPC3-targeting module (HN3) in the tumor and major organs. ( F ) Representative images of healthy and tumor-bearing lungs. ( G ) Particle populations (normalized to the plasma concentration) of the ITGB1 − mscIL12 + HN3 + Deg EVs in the major organs of healthy (Control) or lung metastasized (pulmonary tumor) mice. ** P < 0.01; *** P < 0.001.

Article Snippet: To estimate the loading efficiency of mscIL12 into EVs, reference standard mouse IL12 protein (mIL12, SinoBiological, China) of known quantity and EVs of known number of particles were analyzed by LC/MS, according to a process described elsewhere.

Techniques: Binding Assay, Fluorescence, Incubation, Labeling, Staining, Luciferase, Concentration Assay

Journal: International Journal of Nanomedicine

Article Title: Surface Engineering of HEK293 Cell-Derived Extracellular Vesicles for Improved Pharmacokinetic Profile and Targeted Delivery of IL-12 for the Treatment of Hepatocellular Carcinoma

doi: 10.2147/IJN.S388916

Figure Lengend Snippet: Comparative effects of ITGB1 KO and recombinant mouse IL12 (rmIL12) protein on the anti-tumor efficacy of EVs. ( A ) Tumor growth curves of mice bearing subcutaneous tumors of Hepa1-6-hGPC3 cells, which were treated with intravenous doses of rmIL12 (70ng or 3μg per animal), control EVs (ITGB1 − Deg) or the ITGB1 − mscIL12 + HN3 + Deg EVs (5.0×10 10 p/animal). *,**** P < 0.05, 0.0001 vs ITGB1 − mscIL12 + HN3 + Deg EVs group. ( B ) Tumor size shown after treatment with intravenous doses of rmIL12 (70ng or 3μg per animal), control EVs (ITGB1 − Deg) or the ITGB1 − mscIL12 + HN3 + Deg EVs. ( C ) Heat map of mouse plasma cytokine levels normalized to the control group at 48h post-second dosing (Day 9). ( D ) Flow cytometry analysis of mouse plasma cells for total CD45 + /CD4 + T-cells and CD8 + T-cells 48h after the second dosing (Day 9). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: To estimate the loading efficiency of mscIL12 into EVs, reference standard mouse IL12 protein (mIL12, SinoBiological, China) of known quantity and EVs of known number of particles were analyzed by LC/MS, according to a process described elsewhere.

Techniques: Recombinant, Flow Cytometry