Journal: Journal of experimental and theoretical analyses

Article Title: The Application of Fluorescence Anisotropy for Viscosity Measurements of Small Volume Biological Analytes

doi: 10.3390/jeta1020007

Figure Lengend Snippet: Time-resolved anisotropy measurements of EGFP in different sample volume sizes. Rotational correlation times were derived from time-resolved fluorescence anisotropy measurements of EGFP at different viscosities. The measurements collected with two different analyte volumes (12 μL and 100 μL); ns—not significant; n = 3–4; error bars illustrate standard deviation (SD).

Article Snippet: The fluorescent dyes used in this study were fluorescein disodium salt from Alfa Aesar (Ward Hill, MA, USA) and recombinant EGFP from Novus Biological (NBP1–99915, Centennial, CO, USA) with N- and C-terminal His tags.

Techniques: Derivative Assay, Fluorescence, Standard Deviation

Journal: Journal of experimental and theoretical analyses

Article Title: The Application of Fluorescence Anisotropy for Viscosity Measurements of Small Volume Biological Analytes

doi: 10.3390/jeta1020007

Figure Lengend Snippet: Time-resolved anisotropy measurements of fluorescein end EGFP in 100 μL samples 1.02 cP viscosity standard. Rotational correlation times are specific to the experimental probe used; n = 3–4; error bars illustrate SD.

Article Snippet: The fluorescent dyes used in this study were fluorescein disodium salt from Alfa Aesar (Ward Hill, MA, USA) and recombinant EGFP from Novus Biological (NBP1–99915, Centennial, CO, USA) with N- and C-terminal His tags.

Techniques: Viscosity

Journal: Journal of experimental and theoretical analyses

Article Title: The Application of Fluorescence Anisotropy for Viscosity Measurements of Small Volume Biological Analytes

doi: 10.3390/jeta1020007

Figure Lengend Snippet: The response of EGFP rotational correlation times in a biological sample mimetic. ( A )—comparison of rotational correlation times in samples with or without FBS; ( B )—evaluation of freeze–thaw effects rotational correlation times in samples with FBS. A two-way ANOVA with Sidak post-test was used to compare the data. ns—not significant; n = 3–4; error bars illustrate SD.

Article Snippet: The fluorescent dyes used in this study were fluorescein disodium salt from Alfa Aesar (Ward Hill, MA, USA) and recombinant EGFP from Novus Biological (NBP1–99915, Centennial, CO, USA) with N- and C-terminal His tags.

Techniques: Comparison

Journal: ACS nano

Article Title: Mucosal Immunization with a pH-Responsive Nanoparticle Vaccine Induces Protective CD8 + Lung-Resident Memory T Cells

doi: 10.1021/acsnano.9b00326

Figure Lengend Snippet: (A) Schematic of the experimental timeline. (B) Flow cytometry was used to identify antigen-specific (Tet+) CD8+ T cells in distinct lung compartments (airway, AW; interstitium, IST; marginated vascular, MV) and the spleen. BAL was collected to discriminate AW vs. IST cells, and i.v. staining with αCD45 antibody discriminated IST (CD45−) vs. MV (CD45+) cells. Samples were stained with PE-labeled SIINFEKL/MHC-I tetramer to identify antigen-specific CD8+ T cells. After gating out CD11b+, CD11c+, B220+, and CD4+ cells (“dump”), CD8α+CD45−Tet+ events in AW and IST, CD8α+CD45+Tet+ events in MV, and CD8α+Tet+ events in the spleen were quantified. Dot plots are representative of the gating strategy used in multiple experiments (see Figures S3A–S3C). (C) In conjunction with i.v. staining, microscopy was used to visualize fluorescent OVA-NP conjugates in the lower airways 24 h after immunization. Lungs were stained with αCD45 antibody, which labels vascular leukocytes, and tomato lectin, which binds to capillary endothelial cells and allows for visualization of lung structure. Purple: OVA-NP; blue: vascular leukocytes; green: lung vasculature. Scale bar = 100 μm. Immunization dose: 25 μg NP, 7.5 μg OVA.

Article Snippet: Recombinant vaccinia virus expressing influenza virus nucleoprotein, ovalbumin SIINFEKL peptide, and enhanced green fluorescent protein (VV.NP-S-EGFP) was obtained through the NIH Biodefense and Emerging Infections Research Resources Repository, NIAID, NIH (NR-624; BEI Resources, Manassas, VA).

Techniques: Flow Cytometry, Staining, Labeling, Microscopy

Journal: Journal of Bioscience and Bioengineering

Article Title: Rapid and reliable hybridoma screening method that is suitable for production of functional structure-recognizing monoclonal antibody

doi: 10.1016/j.jbiosc.2021.02.006

Figure Lengend Snippet: Concept of the rMIHS method. (A) After purification of HAT-selected hybridoma, recombinant His-EGFP fusion protein is added into the culture medium. The recombinant protein binds to mIg expressed on the hybridoma cell surface. (B) An example of fluorescent labeling. His-EGFP recombinant protein was specifically labeled to anti-EGFP hybridoma (2H12), but not to anti-αTubulin hybridoma (3A1). White arrows indicate EGFP labelling and nuclei are stained by DAPI. The cells were photographed under an Axiophot 2 fluorescence microscope (Zeiss, Oberkochen, Germany).

Article Snippet: His-EGFP recombinant protein (2 nmol) was mixed with hybridoma in 1 mL RPMI 1640 medium supplemented with 10% Newborn Calf Serum (Biowest, Nuaillé, France), and the cells were incubated for 2 h at 37 °C in 5% CO 2 .

Techniques: Purification, Recombinant, Labeling, Staining, Microscopy

Journal: Journal of Bioscience and Bioengineering

Article Title: Rapid and reliable hybridoma screening method that is suitable for production of functional structure-recognizing monoclonal antibody

doi: 10.1016/j.jbiosc.2021.02.006

Figure Lengend Snippet: Summary of hybridoma generations of rMIHS and ELISA method.

Article Snippet: His-EGFP recombinant protein (2 nmol) was mixed with hybridoma in 1 mL RPMI 1640 medium supplemented with 10% Newborn Calf Serum (Biowest, Nuaillé, France), and the cells were incubated for 2 h at 37 °C in 5% CO 2 .

Techniques: Enzyme-linked Immunosorbent Assay, Clone Assay, Western Blot