Mouse Galectin-9 Antibody

Detection of Galectin-9 in Mouse Thymocytes by Flow Cytometry.

Mouse thymocytes were stained with Goat Anti-Mouse Galectin-9 Short Isoform Affinity-purified Polyclonal Antibody (Catalog # AF3535, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Galectin‑9 in Mouse Splenocytes.

Galectin-9 was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse Galectin-9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3535) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Galectin-9 by Flow Cytometry

Galectin-9 is bound to the surface of primary naive B cells. a Representative flow cytometry plot (left) and quantification (right) of geometric mean ± SEM of surface staining for galectin-9 in WT (black) and Gal9-KO (blue) B cells from nine independent experiments. b Representative DIC (left) and confocal microscopy images (right) mapped to an 8-bit fire color scale (ImageJ) of primary WT (top) and Gal9-KO B cells (bottom) stained for surface galectin-9. Quantification of number of galectin-9 puncta is shown on the right (each dot represents 1 cell, 20 cells measured per condition) with the mean ± SEM indicated by the red bar. Scale bar 2 μm. Data representative of three independent experiments. c Representative confocal microscopy images of cryosections of the inguinal lymph node of WT B cells stained for subcapsular sinus macrophages (CD169; blue), B cells (B220; magenta), and Gal9 (green). Scale bar 20 μm. Data representative of three independent experiments. Statistical significance was assessed by Mann-Whitney, ****p < 0.0001 Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41467-018-05771-8), licensed under a CC-BY license. Not internally tested by R&D Systems.