Journal: International Journal of Molecular Sciences

Article Title: Annexins A2, A6 and Fetuin-A Affect the Process of Mineralization in Vesicles Derived from Human Osteoblastic hFOB 1.19 and Osteosarcoma Saos-2 Cells

doi: 10.3390/ijms22083993

Figure Lengend Snippet: Co-localization of AnxA6, AnxA2, or fetuin-A (FetuA) with TNAP in hFOB 1.19 ( A , C , E ) and Saos-2 ( B , D , F ) cells in resting conditions (R) or after seven-day stimulation with AA and β-GP (S). The cells were incubated with appropriate antibodies: mouse monoclonal anti-AnxA6 ( A , B ), mouse monoclonal anti-AnxA2 ( C , D ), or mouse monoclonal anti-FetuA ( E , F ), all followed by goat anti-mouse IgG-FITC (green); rabbit polyclonal anti-TNAP followed by goat anti-rabbit IgG-TRITC (red) and DAPI for nuclei (blue) and observed under an Axio Observer.Z1 FM (Carl Zeiss, Poznan, Poland) with Phase contrast and appropriate fluorescent filters, magnification 630 x. Arrowheads indicate protein accumulation in vesicular and/or cluster structures. The yellow color and arrows on the merge images indicate AnxA6 ( A , B ), AnxA2 ( C , D ), or FetuA ( E , F ) co-localization with TNAP. Results of a typical experiment are presented.

Article Snippet: Then, cells were incubated with mouse monoclonal anti-annexin A6 (AnxA6; 1:100, v/v ; BD Transduction Laboratories, Warsaw, Poland), mouse monoclonal anti-annexin A2 (AnxA2; 1:100, v/v ; BD Transduction Laboratories, Warsaw, Poland), mouse monoclonal anti-fetuin-A (FetuA; 1:100, v/v ; Abcam, Cambridge, UK), or rabbit polyclonal anti-TNAP (TNAP; 1:100, v/v ; Abcam, Cambridge, UK) primary antibodies diluted in TBS containing 0.5% FBS ( v/v ) and 0.05% Tween-20 ( v/v ).

Techniques: Incubation

Journal: Diagnostics

Article Title: Comparison of 1-Ethyl-3-(3-Dimethylaminopropyl) Carbodiimide Based Strategies to Crosslink Antibodies on Amine-Functionalized Platforms for Immunodiagnostic Applications

doi: 10.3390/diagnostics2030023

Figure Lengend Snippet: ( a ) Surface plasmon resonance (SPR)-based anti-human fetuin A (HFA) immunoassay using various EDC-based strategies for crosslinking anti-HFA antibodies to APTES-functionalized SPR gold chips; ( b ) SPR response units corresponding to the binding of capture anti-HFA antibodies by the various strategies. The values are average of four repeats in different flow cells. The errors bars represent standard deviation.

Article Snippet: The human Fetuin A/AHSG kit with all the necessary components was obtained from R&D Systems Inc., USA.

Techniques: SPR Assay, Binding Assay, Standard Deviation