Journal: Virology

Article Title: Epitope mapping and characterization of a novel CD4-induced human monoclonal antibody capable of neutralizing primary HIV-1 strains

doi: 10.1016/S0042-6822(03)00521-X

Figure Lengend Snippet: Fig. 1. CD4 induction of the binding of the E51 antibody to HIV-1 gp120. The YU2 HIV-1 gp120 glycoprotein was captured on an ELISA plate using the D7324 antibody, which is directed against the gp120 carboxyl-termi- nus. The binding of the 17b and E51 antibodies to the captured gp120 glycoprotein was studied in the absence (open circles) or presence (filled circles) of sCD4 (8 g/ml).

Article Snippet: The ELISA plate was coated with sheep antibody D7324 (Aalto BioReagents, Dublin, Ireland), which recognizes the gp120 carboxylterminus.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: The von Willebrand factor stamps plasmatic extracellular vesicles from glioblastoma patients

doi: 10.1038/s41598-021-02254-7

Figure Lengend Snippet: VWF is transported in circulating EVs from GBM patients. ( A ) Equal volume of protein lysates from EV fractions, obtained with SEC followed by ultracentrifugation, and their corresponding plasma, from one GBM patient and one healthy donor, were separated by SDS-PAGE and analyzed by immunoblot for VWF, FCN3, and HSP70. Blots are representative of n = 3. ( B ) mRNA expression of von Willebrand factor (VWF) and ficolin-3 (FCN3) in healthy subjects and GBM patients estimated from The Cancer Genome Atlas (TCGA, n > 500 for GBM patients). ( C ) Anomalies currently reported in VWF and FCN3 genes. ( D ) Equal volume of protein lysates from EV fractions (3 GBM and 3 Healthy) were separated by SDS-PAGE and analyzed by immunoblot for VWF, GM130 (putative intracellular membrane protein contaminant), and Apolipoprotein A1 (APOA1, plasmatic protein). Healthy donor plasma and total cell lysate from GBM cells serve as controls. Blots are representative of n > 7 individual samples. ( E ) Densitometric analysis was performed on 7 and 8 independent EV preparations from Healthy and GBM liquid biopsies, respectively. ( F ) Concentration of VWF in plasmatic EVs from healthy donors and GBM patients (n = 20). Mann–Whitney test, * p < 0.05, ** p < 0.01, *** p < 0.001. S.D. are shown in panel E and S.E.M. in panel F.

Article Snippet: EVs were separated and concentrated from 500 μl of plasma as described above and the pellet was directly lysed in boiling Laemmli for 10 min. Proteins were resolved by SDS-PAGE, transferred onto nitrocellulose membranes and blotted with the following antibodies, all diluted at 1/1000: CD9 (EXOAB-CD9A-1, SBI), CD63 (EXOAB-CD63A-1, SBI), VWF (sc-53466, Santa Cruz), FCN3 (CL7767AP, Cedarlane) , Apolipoprotein A1 (sc-376818, Santa Cruz), HSP70 (EXOAB-HSP70A, SBI), and GM130 (ab52649, Abcam) diluted at 1/1000.

Techniques: Clinical Proteomics, SDS Page, Western Blot, Expressing, Membrane, Concentration Assay, MANN-WHITNEY