Journal: Scientific Reports

Article Title: A fixation method for the optimisation of western blotting

doi: 10.1038/s41598-019-43039-3

Figure Lengend Snippet: Fixation-dependent differences in lectin staining intensities when using PVDF and nitrocellulose membranes. Pooled human serum proteins (3 μg) were separated on 10% SDS-PAGE, and the proteins were transferred onto PVDF and nitrocellulose membranes, the whole membrane was cutted into five pieces for subsequently fixation treatments and followed by staining with lectins (LCA and SNA). Lane i, CBB staining; lane ii, no fixation; lane iii, drying at room temperature; lane iv, sample heating at 100 °C; lane v, organic solvent (acetone and 50% methanol for PVDF and nitrocellulose membranes, respectively) treatments at room temperature; lane vi, organic solvent treatments followed by sample heating at 100 °C. All treatments were applied for 30 min. Left, WB pattern; right, quantitative analysis (n = 3 individual experiments). The exposure times were the same in all procedures. Band intensities were analysed and compared using Image Lab software (Bio-Rad Laboratories) and GraphPad Prism version 6. ** Significantly different p < 0.01, *** p < 0.001, **** p < 0.0001. All values are means ± S.E. (error bars).

Article Snippet: Biotinylated lectins, LCA, SNA, PHA-E, PHA-L, and AAL (Supplementary Table ), were purchased from Vector Laboratories Inc. (Burlingame, CA, USA).

Techniques: Staining, SDS Page, Software

Journal: Analytical and bioanalytical chemistry

Article Title: Heparin-immobilized microspheres for the capture of cytokines

doi: 10.1007/s00216-010-4170-1

Figure Lengend Snippet: Effect of heparin loading on the binding capacity of heparin-immobilized microspheres to human aFGF (30 ng/mL). The fluorescence signal on the beads was developed by incubating the beads with PE-labeled antibody and measured on the BD FACSArray flow cytometer. The MFI values were significantly different for the microspheres with varying heparin loading amounts at the 95% confidence level (one-way ANOVA). The error bars represent the standard deviations of the mean intensities based on three independent experiments.

Article Snippet: Recombinant human aFGF and human aFGF ELISA kit (biotinylated rabbit anti-human aFGF polyclonal antibody included) were obtained from Antigenix America, Inc. (Huntington Station, NY, USA).

Techniques: Binding Assay, Fluorescence, Labeling, Flow Cytometry

Journal: Analytical and bioanalytical chemistry

Article Title: Heparin-immobilized microspheres for the capture of cytokines

doi: 10.1007/s00216-010-4170-1

Figure Lengend Snippet: Effect of free heparin on the interaction between the cytokine, CCL2 (■) or aFGF (▲), and the heparin-immobilized microspheres. The fluorescence signal on the beads was developed by incubating the beads with corresponding PE-labeled antibody and measured on the BD FACSArray flow cytometer. The error bars represent the standard deviations of the mean intensities based on three independent experiments.

Article Snippet: Recombinant human aFGF and human aFGF ELISA kit (biotinylated rabbit anti-human aFGF polyclonal antibody included) were obtained from Antigenix America, Inc. (Huntington Station, NY, USA).

Techniques: Fluorescence, Labeling, Flow Cytometry

Journal: Analytical and bioanalytical chemistry

Article Title: Heparin-immobilized microspheres for the capture of cytokines

doi: 10.1007/s00216-010-4170-1

Figure Lengend Snippet: Binding of cytokines, (a) aFGF, (b) CCL2, (c) VEGF, and (d) CCL5, onto the heparin-immobilized microspheres (■) and the acetylated amino microspheres (□). The fluorescence signal on the beads was developed by incubating the beads with corresponding PE-labeled antibody and measured on the BD FACSArray flow cytometer (aFGF and CCL2) or the Luminex system (VEGF and CCL5). Inset: binding of each cytokine to the heparin-immobilized microspheres with background binding to the acetylated amino microspheres subtracted. The error bars represent the standard deviations of the mean intensities based on three independent experiments.

Article Snippet: Recombinant human aFGF and human aFGF ELISA kit (biotinylated rabbit anti-human aFGF polyclonal antibody included) were obtained from Antigenix America, Inc. (Huntington Station, NY, USA).

Techniques: Binding Assay, Fluorescence, Labeling, Flow Cytometry, Luminex

Journal: Analytical and bioanalytical chemistry

Article Title: Heparin-immobilized microspheres for the capture of cytokines

doi: 10.1007/s00216-010-4170-1

Figure Lengend Snippet: Binding kinetics of cytokines, aFGF (■), VEGF (●), CCL2 (▲), and CCL5 (▼), onto the heparin-immobilized microspheres. % Bound cytokine was calculated as the mass ratio of captured cytokine to the total cytokine initially added. The error bars represent the standard deviations of the mean values based on three independent experiments.

Article Snippet: Recombinant human aFGF and human aFGF ELISA kit (biotinylated rabbit anti-human aFGF polyclonal antibody included) were obtained from Antigenix America, Inc. (Huntington Station, NY, USA).

Techniques: Binding Assay