Journal: Toxins

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59

doi: 10.3390/toxins7010110

Figure Lengend Snippet: Quantitative measurement ( K d ) of the affinity of anti-VLY monoclonal antibodies (MAbs) with vaginolysin (VLY) mutants.

Article Snippet: Plates were blocked for 1 h at RT with 1% BSA (PAA Laboratories GmbH, Pasching, Austria) in PBS and then incubated with 100 μL serially diluted anti-VLY MAbs 9B4 and 21A5 (concentration ranged from 1.9 × 10 −7 to 3.3 × 10 −12 M in PBS with 0.1% Tween-20 (Carl Roth, Karlsruhe, Germany)) for 1 h at RT.

Techniques:

Journal: Toxins

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59

doi: 10.3390/toxins7010110

Figure Lengend Snippet: Binding of native vaginolysin (VLY) to CHO and CHO-hCD59 cells probed with monoclonal antibodies (MAbs) 9B4 and 21A5 detected by flow cytometry. The figures are representative of experiments performed at least in triplicate. GMF: geometric mean fluorescence.

Article Snippet: Plates were blocked for 1 h at RT with 1% BSA (PAA Laboratories GmbH, Pasching, Austria) in PBS and then incubated with 100 μL serially diluted anti-VLY MAbs 9B4 and 21A5 (concentration ranged from 1.9 × 10 −7 to 3.3 × 10 −12 M in PBS with 0.1% Tween-20 (Carl Roth, Karlsruhe, Germany)) for 1 h at RT.

Techniques: Binding Assay, Flow Cytometry, Fluorescence

Journal: Toxins

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59

doi: 10.3390/toxins7010110

Figure Lengend Snippet: ( A ) The overlay histograms represent binding profile of vaginolysin mutant VLY(ML) to CHO and CHO-hCD59 cells probed with monoclonal antibody (MAb) 9B4; ( B ) Binding of mutant protein VLY(ML) to CHO and CHO-hCD59 cells probed with MAbs 9B4 and 21A5 detected by flow cytometry. The figures are representative of experiments performed at least in triplicate. GMF: geometric mean fluorescence.

Article Snippet: Plates were blocked for 1 h at RT with 1% BSA (PAA Laboratories GmbH, Pasching, Austria) in PBS and then incubated with 100 μL serially diluted anti-VLY MAbs 9B4 and 21A5 (concentration ranged from 1.9 × 10 −7 to 3.3 × 10 −12 M in PBS with 0.1% Tween-20 (Carl Roth, Karlsruhe, Germany)) for 1 h at RT.

Techniques: Binding Assay, Mutagenesis, Flow Cytometry, Fluorescence

Journal: Toxins

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59

doi: 10.3390/toxins7010110

Figure Lengend Snippet: ( A ) Binding of vaginolysin mutant protein VLY(ML) preincubated with cholesterol to CHO and CHO-hCD59 cells probed with monoclonal antibodies (MAbs) 9B4 and 21A5; ( B ) Binding of VLY-TL mutant with impaired cholesterol recognition motif to CHO cells detected with 9B4 and 21A5 MAbs. The figures are representative of experiments performed at least in triplicate. GMF: geometric mean fluorescence.

Article Snippet: Plates were blocked for 1 h at RT with 1% BSA (PAA Laboratories GmbH, Pasching, Austria) in PBS and then incubated with 100 μL serially diluted anti-VLY MAbs 9B4 and 21A5 (concentration ranged from 1.9 × 10 −7 to 3.3 × 10 −12 M in PBS with 0.1% Tween-20 (Carl Roth, Karlsruhe, Germany)) for 1 h at RT.

Techniques: Binding Assay, Mutagenesis, Fluorescence