biotinylated anti human mgl antibody  (R&D Systems)

Journal: bioRxiv

Article Title: CLR-Seq: a pipeline to identify bacterial microbiota species with immunomodulatory potential through human C-type lectin receptor interaction

doi: 10.1101/2025.03.31.646320

Figure Lengend Snippet: Isolated fecal microbiota were supplemented with 10 6 CFU langerin-reactive S. aureus N315 (upper panels) or MGL-reactive PS187 (lower panels). S. aureus strains were fluorescently labeled with CellTrace Violet prior to addition to the microbiota suspension. For both langerin and MGL, the bacteria were measured without (left panels) and after FITC-labeled CLR staining (right panel). The reduced relative abundance of CellTrace Violet+/MGL+ S. aureus results from agglutination by the secondary antibody. The percentages are shown per quadrant. The images shown are representative of two different fecal donors.

Article Snippet: To determine CLR-specific binding of fecal bacteria, the filtered microbiota suspension was centrifuged and stained with 25 µg/mL recombinant human langerin-FITC [ ] or his-tagged human MGL (hMGL; R&D Systems) in Tris-sodium-magnesium buffer (20 mM Tris; Roche, 150 mM NaCl; Merck, 2 mM CaCl2; Merck, 2 mM MgCl2; Merck, pH 7.0; TSM) + 1% BSA, and incubated 30 minutes at 37°C with agitation (600 rpm).

Techniques: Isolation, Labeling, Suspension, Bacteria, Staining, Agglutination

Journal: bioRxiv

Article Title: CLR-Seq: a pipeline to identify bacterial microbiota species with immunomodulatory potential through human C-type lectin receptor interaction

doi: 10.1101/2025.03.31.646320

Figure Lengend Snippet: Correlation of the average IgA enrichment scores for each genus with their respective A. langerin (n=93) or B. MGL (n=94) average enrichment scores. All genera shown were present in the IgA+ and CLR+ samples of all individual donors. Spearman’s rank order correlation was used for statistical analysis. Monoderm genera are shown in orange, diderm genera in purple. The ten genera with the highest and lowest probability scores are shown for C. langerin or D. MGL binding. Individual probability are shown for each donor. A dotted line is shown to mark the probability score at which a genus would have equal relative abundance in the unsorted and sorted samples for the respective sorting condition. Diderm genera are underscored.

Article Snippet: To determine CLR-specific binding of fecal bacteria, the filtered microbiota suspension was centrifuged and stained with 25 µg/mL recombinant human langerin-FITC [ ] or his-tagged human MGL (hMGL; R&D Systems) in Tris-sodium-magnesium buffer (20 mM Tris; Roche, 150 mM NaCl; Merck, 2 mM CaCl2; Merck, 2 mM MgCl2; Merck, pH 7.0; TSM) + 1% BSA, and incubated 30 minutes at 37°C with agitation (600 rpm).

Techniques: Binding Assay

Journal: bioRxiv

Article Title: CLR-Seq: a pipeline to identify bacterial microbiota species with immunomodulatory potential through human C-type lectin receptor interaction

doi: 10.1101/2025.03.31.646320

Figure Lengend Snippet: Binding of A. recombinant human langerin-FITC and B. recombinant human MGL detected by an anti-hisTag-FITC antibody to S. aureus positive controls and identified microbiota species. CLRs were used at the same concentration (25 µg/mL) as used for fluorescence cell sorting. Data are shown as the mean of the geometric mean fluorescence intensity (FI) ± standard error of mean from three independent experiments. **** P < .0001

Article Snippet: To determine CLR-specific binding of fecal bacteria, the filtered microbiota suspension was centrifuged and stained with 25 µg/mL recombinant human langerin-FITC [ ] or his-tagged human MGL (hMGL; R&D Systems) in Tris-sodium-magnesium buffer (20 mM Tris; Roche, 150 mM NaCl; Merck, 2 mM CaCl2; Merck, 2 mM MgCl2; Merck, pH 7.0; TSM) + 1% BSA, and incubated 30 minutes at 37°C with agitation (600 rpm).

Techniques: Binding Assay, Recombinant, Concentration Assay, Fluorescence, FACS