biotinylated goat anti mouse c3d  (R&D Systems)

Journal: The Journal of Clinical Investigation

Article Title: Impaired complement regulation drives chronic lung allograft dysfunction after lung transplantation

doi: 10.1172/JCI188891

Figure Lengend Snippet: ( A ) Levels of C3d bound to B cells 2 days after the induction of DOX-induced allograft club cell injury. Representative FACS histogram of C3d B cell staining and (right panel) plots of C3d MFI normalized to a corresponding fluorescence minus one (FMO) control ( n = 4/group). ( B ) POD 16 FACS contour plot of intragraft B cell abundance (left), ki67 histogram (proliferation) (middle), and a plot of total allograft B cell numbers on POD 16 (right). FACS contour plots and histograms are representative results from 4/group. ( C ) Percentages of BAL CD19 + lymphocytes in UCSF cohorts who carry the GG genotype and C3 R102G (CC&GC) polymorphisms. P = 0.02 by an unpaired t test. ( D ) Quantitation of BAL B cell numbers in POD 16 allografts ( n = 4/group). Levels of POD 16 of ( E ) BAL ( n = 4/group) and ( F ) serum ( n = 5/group) donor reactive IgM, IgG 1 , IgG 2c , and IgG 3 . ( G ) POD 16, intragraft CD138 + IgM + and IgG + Ab-secreting cell numbers ( n = 5/group). ( H ) A representative FACS gating strategy used to identify intragraft CD80 + CD73 + CD273 + memory B cells, as shown in the bar graphs ( I ), which display the total intragraft numbers of these cells expressing either surface IgM ( n = 5/group) or IgG ( n = 4/group) on POD 16. Bar graphs and dot blots show mean ± SD values according to 2-way ANOVA with Šidák’s multiple comparisons test ( A ), and Welch’s t test ( B , D – G , and I ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cell surface staining was performed with the following Abs: CD45 (clone 30-F11; eBioscience), CD45.2 (clone 104; BioLegend), CD90.2 (clone 53-2.1; eBioscience), CD4 (clone RM4-5; eBioscience), CD8α (clone 53-6.7; eBioscience), B220 (RA3-6B2; Biolegend), CD19 (1D3; BD Biosciences), rabbit anti–human/mouse/rat C3d (catalog bs-4877R; Bioss), CD138 (281-2; Biolegend) CD38 (S21016F; Biolegend), IgD (11-26c.2a; Biolegend), Foxp3 (FJK-16s; Thermo Fisher), CD73 (TY/11.8; Biolegend), CD80 (16-10A1; Biolegend), CD273 (TY25; Thermo Fisher), CD31 (clone 390; BioLegend), CD34 (clone HM34; BioLegend), and CD326 (clone G8.8; BioLegend).

Techniques: Staining, Fluorescence, Control, Quantitation Assay, Expressing

Journal: The Journal of Clinical Investigation

Article Title: Impaired complement regulation drives chronic lung allograft dysfunction after lung transplantation

doi: 10.1172/JCI188891

Figure Lengend Snippet: C3d and CD19 immunofluorescent staining of Crry –/– ( A ) and wild-type Crry +/+ ( B ) recipient allograft tissue (LTx). Stars and arrows denote C3d staining on vascular endothelium and bronchial epithelium, respectively. Rectangular insets depict tertiary lymphoid aggregates enriched with B cells. Scale bars: 150 μm. The images shown are representative results of at least 3 transplants per group.

Article Snippet: Cell surface staining was performed with the following Abs: CD45 (clone 30-F11; eBioscience), CD45.2 (clone 104; BioLegend), CD90.2 (clone 53-2.1; eBioscience), CD4 (clone RM4-5; eBioscience), CD8α (clone 53-6.7; eBioscience), B220 (RA3-6B2; Biolegend), CD19 (1D3; BD Biosciences), rabbit anti–human/mouse/rat C3d (catalog bs-4877R; Bioss), CD138 (281-2; Biolegend) CD38 (S21016F; Biolegend), IgD (11-26c.2a; Biolegend), Foxp3 (FJK-16s; Thermo Fisher), CD73 (TY/11.8; Biolegend), CD80 (16-10A1; Biolegend), CD273 (TY25; Thermo Fisher), CD31 (clone 390; BioLegend), CD34 (clone HM34; BioLegend), and CD326 (clone G8.8; BioLegend).

Techniques: Staining