monoclonal mouse anti human c5b 9 antibody  (Hycult Biotech)

Journal: Medical Microbiology and Immunology

Article Title: Lipopolysaccharide with long O-antigen is crucial for Salmonella Enteritidis to evade complement activity and to facilitate bacterial survival in vivo in the Galleria mellonella infection model

doi: 10.1007/s00430-024-00790-3

Figure Lengend Snippet: C5b-9 deposition and the effect of complement attack on the cell membrane integrity of S . Enteritidis PCM 2817 WT and mutant strains. A Mean fluorescence intensity (MFI) of C5b-9 neo-epitope deposition on bacterial cells. B Frequency of 7AAD positive cells; the percentage of 7AAD positive cells stands for the percentage of cells with compromised membrane integrity. The experiment was performed in three biological replicates. Data was analyzed by ordinary one-way ANOVA and Dunnett's post hoc test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001), assuming WT as the control group. Average from six measurements ± SD

Article Snippet: Cells were incubated with the monoclonal mouse anti-human C5b-9 antibody (clone aE11) (Hycult) for 45 min at ambient temperature, washed and detected with an anti-mouse Alexa 488 antibody (Abcam) (45 min, ambient temperature).

Techniques: Membrane, Mutagenesis, Fluorescence, Control

Journal: Frontiers in Immunology

Article Title: Targeted complement inhibition using bispecific antibodies that bind local antigens and endogenous complement regulators

doi: 10.3389/fimmu.2024.1288597

Figure Lengend Snippet: Targeted bispecific antibodies that bind endogenous complement inhibitors decrease complement activation in situ. BsAbs were functionally tested in plate-bound complement activation assays. Plates coated with BSA-DNP and IgG (classical pathway), acetylated HSA (lectin pathway), or LPS (alternative pathway) were incubated with a titration of anti-FH x anti-DNP (A, C) , 5 µg/mL anti-FH x anti-DNP (B, D) , a titration of anti-C4BP x anti-DNP (E, G) , or 5 µg/mL anti-C4BP x anti-DNP (F, H) or control bsAbs, with NHS with (+) or without (-) exogenous FH (A–D) or C4BP (E–H) . FH (A, B) , C4BP (E, F) , and C5b-9 (C, D, G, H) were detected. In (A, C, E, G) IgG-coated plates were used. Bars indicate means and error bars indicate standard deviations of one representative experiment out of at least three experiments. One-way ANOVA compared to no bsAb, **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: Next, FH and C4BP were detected as described above for the bispecificity ELISA (using either goat anti-human FH and HRP-labeled rabbit anti-goat Ig, or using rabbit anti-human C4BP and HRP-labeled goat anti-rabbit Ig), and C5b-9 was detected using mouse anti-human C5b-9 (Dako, M0777) and HRP-labeled goat anti-mouse Ig (Dako, P0447), all diluted in PTB buffer.

Techniques: Activation Assay, In Situ, Incubation, Titration, Control