Mouse Dectin-1/CLEC7A Antibody

Detection of Yeast Dectin-1/CLEC7A by Immunocytochemistry/Immunofluorescence Binding of cHis-sDectin-1 to Saccharomyces cerevisiae cells.Purified cHis-sDectin-1 was diluted to a concentration of 25 µg/ml in blocking buffer with 500 µg/ml, 100 µg/ml or no laminarin. This was added to Saccharomyces cerevisiae yeast cells from overnight culture in blocking buffer (PBS with 3% FBS). The cells were then probed with an mDectin-1 goat polyclonal antibody (1∶200; AF1756; R&D Systems) and AlexaFluor546-conjugated anti-goat antibody (1∶100; Catalog Number A-11056; Molecular Probes). After which, the cells were washed with PBS and fixed using 4% paraformaldehyde. The cells were then resuspended in PBS and visualized by phase contrast and fluorescent microscopy at (A) 200× magnification and (B) 600× magnification. Images are cropped or scaled to fit the illustration. cHis-sDectin-1 stained yeast cells was also analyzed by flow cytometry (C). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0052785), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Yeast Dectin-1/CLEC7A by Immunocytochemistry/Immunofluorescence Binding of cHis-sDectin-1 to Saccharomyces cerevisiae cells.Purified cHis-sDectin-1 was diluted to a concentration of 25 µg/ml in blocking buffer with 500 µg/ml, 100 µg/ml or no laminarin. This was added to Saccharomyces cerevisiae yeast cells from overnight culture in blocking buffer (PBS with 3% FBS). The cells were then probed with an mDectin-1 goat polyclonal antibody (1∶200; AF1756; R&D Systems) and AlexaFluor546-conjugated anti-goat antibody (1∶100; Catalog Number A-11056; Molecular Probes). After which, the cells were washed with PBS and fixed using 4% paraformaldehyde. The cells were then resuspended in PBS and visualized by phase contrast and fluorescent microscopy at (A) 200× magnification and (B) 600× magnification. Images are cropped or scaled to fit the illustration. cHis-sDectin-1 stained yeast cells was also analyzed by flow cytometry (C). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0052785), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Yeast Dectin-1/CLEC7A by Immunocytochemistry/Immunofluorescence Binding of cHis-sDectin-1 to Saccharomyces cerevisiae cells.Purified cHis-sDectin-1 was diluted to a concentration of 25 µg/ml in blocking buffer with 500 µg/ml, 100 µg/ml or no laminarin. This was added to Saccharomyces cerevisiae yeast cells from overnight culture in blocking buffer (PBS with 3% FBS). The cells were then probed with an mDectin-1 goat polyclonal antibody (1∶200; AF1756; R&D Systems) and AlexaFluor546-conjugated anti-goat antibody (1∶100; Catalog Number A-11056; Molecular Probes). After which, the cells were washed with PBS and fixed using 4% paraformaldehyde. The cells were then resuspended in PBS and visualized by phase contrast and fluorescent microscopy at (A) 200× magnification and (B) 600× magnification. Images are cropped or scaled to fit the illustration. cHis-sDectin-1 stained yeast cells was also analyzed by flow cytometry (C). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0052785), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Dectin-1/CLEC7A by Western Blot MTX amplification and characterization of sDectin-1 cell pools.(A) Western blotting of supernatant samples from cultures adapted to different MTX concentrations using an mDectin-1 goat polyclonal antibody (1∶1000; AF1756; R&D Systems) with a HRP conjugated anti-goat antibody (1∶2000; Catalog number V8051; Promega). cHis pool and nHis pool are cell pools producing sDectin-1 with histidine tagged at the C- and N-terminals respectively. (B) Shake flask batch culture cell growth and sDectin-1 production profiles of sDectin-1 producing cell pools. Values shown represent mean values obtained from three replicate flasks. Error bars indicate the standard deviation of the experiment. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0052785), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Mouse Dectin-1/CLEC7A Antibody by Western Blot OatN beta -glucan binds to HPCA, promotes microglia up-take of oatN and inhibits dectin-1 activation, whereas OatN digalactosyldiacylglycerol (DGDG) inhibits oatN beta -glucan access to dectin-1. H) A representative western blot of p-Syk in BV2 cells treated with Eth (200 mm) or Eth plus OBG (20 mg mL−1). Results represent 1 out of 3 independent experiments. Band intensity was quantified and normalized to beta -actin and the results are presented between the panels. I) The surface plasmon resonance (SPR) assay was performed to determine OBG binding to dectin-1. (Upper panel) BG (commercial beta -glucan, 100 ng mL−1), PBLP (liposomes made by peripheral blood cell derived lipids), OatN (20 mg mL−1), and OBG (20 mg mL−1) were used as analytes. (Bottom panel) OBG-DGDG (OBG remove DGDG, 20 or 60 mg mL−1) and OBG-PC (OBG remove PC, 20 mg mL−1) were used as analytes. Results represent 1 out of 3 independent experiments. J) Dectin-1 binding to HPCA-Rab11a complex was detected by immunoprecipitation. HPCA complex in BV2 cells treated with Eth or OBG was pulled down by anti-HPCA antibody (1 mg mL−1). Dectin-1 and Rab11a were probed in a western blot. Results represent 1 out of 3 independent experiments. Band intensity was quantified and normalized by HPCA, and the results are presented between the panels. K) Confocal images of dectin-1 (green) overlay with Rab11a (red) in BV2 cells treated with Eth (200 mm) or Eth plus OBG (20 mg mL−1). Results represent 1 out of 3 independent experiments. Five random fields were photographed, and representative results are shown. Scale bars, 20 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/34897972), licensed under a CC-BY license. Not internally tested by R&D Systems.