Mouse OCILRP2/CLEC2i Antibody

Detection of Mouse OCILRP2/CLEC2i by Immunocytochemistry/Immunofluorescence Membrane co-localization of OCILRP2 and DAP12 under anti-CD3/CD28 mAb treatment in EL4 T cells.EL4 T cells pre-transfected with pEGFP-siOCILRP2 or pre-coated with the OCILRP2 Ab were cultured overnight in the presence or absence of an anti-CD3/CD28 mAb and then stained for OCILRP2 (green) and nuclear stained with DAPI (blue) and DAP12 (red) to study OCILRP2 and DAP12 protein expression and localization. Unstimulated EL4 T cells exhibited OCILRP2 protein expression in the cytoplasm (upper panels). In contrast, CD3/CD28-activated EL4 T cells showed intracellular and membrane OCILRP2. OCILRP2/DAP12 co-localization appears in yellow. Each picture is representative of the vast majority of the observed cells on the slides. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0113218), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse OCILRP2/CLEC2i by Western Blot Interaction of OCILRP2 and DAP12 in GST pull-down and co-immunoprecipitation assays.The GST pull-down assay was carried out using purified beads that contained GST, GST-DAP12, or GST-OCILRP2. Precipitated OCILRP2 or DAP12 was detected by western blotting using an anti-OCILRP2 or anti-DAP12 antibody, respectively (a, b). 293T cells were grown in 6-cm dishes and transfected with the pCDNA3-HA-OCILRP2 and pDsRed-C1-DAP12 plasmids, respectively. OCILRP2 and DAP12 were detected by western blotting using an anti-HA antibody or an anti-DAP12 antibody (c). Schematic diagram of OCILRP2 predicted by SMART software. The green column represents the transmembrane region (amino acids 57–80) (d). The GST pull-down assay was carried out using purified beads that contained GST, full-length GST-OCILRP2, GST-OCILRP2i (aa 1–57), or GST-OCILRP2e (aa 80–221). Precipitated DAP12 was detected by western blotting using an anti-DAP12 antibody (e). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0113218), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse OCILRP2/CLEC2i by Western Blot Interaction of OCILRP2 and DAP12 in GST pull-down and co-immunoprecipitation assays.The GST pull-down assay was carried out using purified beads that contained GST, GST-DAP12, or GST-OCILRP2. Precipitated OCILRP2 or DAP12 was detected by western blotting using an anti-OCILRP2 or anti-DAP12 antibody, respectively (a, b). 293T cells were grown in 6-cm dishes and transfected with the pCDNA3-HA-OCILRP2 and pDsRed-C1-DAP12 plasmids, respectively. OCILRP2 and DAP12 were detected by western blotting using an anti-HA antibody or an anti-DAP12 antibody (c). Schematic diagram of OCILRP2 predicted by SMART software. The green column represents the transmembrane region (amino acids 57–80) (d). The GST pull-down assay was carried out using purified beads that contained GST, full-length GST-OCILRP2, GST-OCILRP2i (aa 1–57), or GST-OCILRP2e (aa 80–221). Precipitated DAP12 was detected by western blotting using an anti-DAP12 antibody (e). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0113218), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse OCILRP2/CLEC2i by Flow Cytometry Membrane re-localization of OCILRP2 via treatment with an anti-CD3/CD28 mAb.EL4 cells were incubated with plate-bound CD3/CD28 antibodies, isotype-matched mIgGs, or PMA/ionomycin for 48 h. The cells (1×106 cells) were then incubated with an anti-OCILRP2 antagonist antibody for 30 min on ice, followed by staining with an FITC-labeled second antibody. The cells were analyzed with Cellquest software using a FACS Calibur flow cytometer. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0113218), licensed under a CC-BY license. Not internally tested by R&D Systems.