rat anti mbl c  (Hycult Biotech)

Journal: Frontiers in Immunology

Article Title: Inhibition of the lectin pathway of complement activation reduces LPS-induced acute respiratory distress syndrome in mice

doi: 10.3389/fimmu.2023.1192767

Figure Lengend Snippet: Binding of the LP recognition molecules and C1q to LPS. Solid-phase binding assays were used to assess the binding of the LP recognition molecules from murine (A, C, E) or human sera (B, D, F) to LPS. High levels of CL-11, mouse MBL-C and human MBL were observed. Neither ficolins nor C1q bound to LPS (G, H) . Results are means of duplicates ± SD.

Article Snippet: Plates were washed and binding of recognition molecules was detected using goat anti-CL-11(Santa-Cruz Biotechnology), rabbit anti-human L-ficolin, mouse anti-human CL-11, rat anti-mouse MBL-A, rat anti-mouse MBL-C, rabbit anti-mouse ficolin-A or rabbit anti-C1q antibodies.

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Human L-ficolin, a Recognition Molecule of the Lectin Activation Pathway of Complement, Activates Complement by Binding to Pneumolysin, the Major Toxin of Streptococcus pneumoniae

doi: 10.1371/journal.pone.0082583

Figure Lengend Snippet: Serial dilutions of NMS or NHS in BBS were incubated in ELISA plates pre-coated with PLY or specific ligands for LP recognition molecules. The binding of specific LP recognition molecules to PLY was subsequently detected using antibodies directed against MBL, ficolins and CL-11. The binding of murine MBL-A and MBL-C (A), murine ficolin-A (C), murine CL-11 (E), human MBL (B), human FCN2 & 3 (D) and human CL-11 (F) were assessed by ELISA. In contrast to mouse ficolin-A (which has no binding affinity to PLY) human L-ficolin (FCN2) (the human orthologoue of murine ficolin-A) showed strong binding to PLY. Results are means (±SEM) of three independent experiments.

Article Snippet: Plates were washed and bound proteins were detected using monoclonal rat anti-mouse MBL-A (Hycult), rat anti-mouse MBL-C (Hycult), rabbit anti-mouse ficolin-A, rabbit anti-human L-ficolin (Sigma), mouse anti-human H-ficolin (Hycult), mouse anti-human CL-11, rat anti-mouse CL-11 mAb, rabbit anti-human C1q.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: bioRxiv

Article Title: Germinal center block exacerbates extrafollicular responses and accelerates autoimmune disease progression in a murine lupus model

doi: 10.1101/2022.03.04.482991

Figure Lengend Snippet: GC block causes increased levels of autoantibodies and PB/PCs in SLE-like mice. ( A ) Schematic overview of experimental setup and treatment protocol. Bcl-6 flx/flx (Cre-, purple) and Aicda-Cre Bcl-6 flx/flx (Cre+, blue) mice were either left untreated (dark color, n = 7 and n = 8, respectively) or treated with R848 (light color, n = 6 and n= 8, respectively), as indicated. ( B ) Spleen weights. ( C ) Anti-dsDNA IgG2c levels. ( D ) Total IgG2c levels. ( E ) Total IgG1 levels. ( F ) Total IgG3 levels. ( G ) Representative confocal micrograph of spleen from a Cre-untreated animal, stained for CD169 (red), IgD (blue) and Ki67 (green). Scale bar is 400 µm ( H ). As G, but for a Cre-R848-treated animal. ( I ) As G, but for a Cre+ untreated animal. ( J ) As G, but for a Cre+ R848-treated animal. ( K ) High-resolution image of a GC from a Cre-R848-treated mouse. Scale bar is 100 µm. L ) GC per follicle in spleen. ( M ) Flow cytometry analyses of monocyte frequencies in IngLN, MesLN, and spleen (Ly6CG int of live, singlet leukocytes). ( N ) As M, but neutrophil frequencies (Ly6CG hi of live, singlet leukocytes). ( O ) As M, but B cell frequencies (B220 + CD4 - CD8 - of live, singlet leukocytes). ( P ) As M, but GCB frequencies (CD95 hi CD38 lo of B cells). ( Q ) As M, but PB and PC frequencies (CD138 hi of live, singlet leukocytes). Data are pooled from two independent experiments. Bar graphs show mean ± SD. Two-way ANOVA with Holm-Sidak’s post hoc test was used to analyze the data. ns = p≥0.05, * = p<0.05, ** = p<0.01, *** = p<0.001.

Article Snippet: In brief, antibody (either “14D12” rat IgG2a to mouse MBL-C (Hycult Biotech), or “RTK2758” rat IgG2a isotype control (Abcam)) was concentrated in antibody preparation buffer to a concentration of 2 mg/mL or above.

Techniques: Blocking Assay, Staining, Flow Cytometry