Human/Mouse/Rat DEP-1/CD148 Antibody

Detection of Human DEP-1/CD148 by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat DEP-1/CD148 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1934) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for DEP-1/CD148 at approximately 220 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Monkey DEP-1/CD148 by Immunocytochemistry/Immunofluorescence

Proximity ligation assay reveals association of DEP-1 with its substrate FLT3.(A) COS7 cells were transiently transfected with expression constructs for FLT3, DEP-1, the catalytically inactive DEP-1 C1239S trapping mutant, or corresponding control plasmids as indicated. Complex formation was measured by in situ PLA using rabbit anti-FLT3 antibodies, goat anti-DEP-1 antibodies, and corresponding secondary reagents. In situ PLA is indicated by red signals of the rolling cycle amplification products (RCPs). FLT3 expression (green) was visualized by additional staining with FITC-labeled anti-rabbit IgG antibodies; nuclei (blue) were counterstained with Hoechst 33342. Scale bars 20 µm. (B), (C) Complex formation of endogenous DEP-1 with endogenous FLT3 in THP-1 cells. Cells were transfected with DEP-1-specific or control siRNA by Nucleofection, were then starved and either left unstimulated or were stimulated with FL (100 ng/ml) for 10 min as indicated. (B) Example images; DEP-1 knockdown efficiency was detected by immunblotting (lower panel). DEP-1-FLT3 complexes as RCPs are shown in red, nuclei are depicted in blue and scale bars represent 20 µm for the overview images and 5 µm for the insets. (C) Quantification of detected in situ PLA signals over 5 images per variant. The data are representative for 3 experiments with consistent results. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/23650535), licensed under a CC-BY license. Not internally tested by R&D Systems.