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.

Detection of DEP-1/CD148 by Western Blot

Rescue of Defective ERK Signaling in VEGF-A-Stimulated Nrp1cyto Arterial EC by Knockdown of PTP1bPrimary arterial EC from Nrp1cyto mice transfected with siRNA specific for the indicated phosphatases were serum-starved and then stimulated with 50 ng/ml VEGF-A165.(A and B) Knockdown of the indicated phosphatases in Nrp1cyto arterial EC shown by immunoblotting (A); PTP1b knockdown was quantified in (B), dashed line indicates normal expression levels.(C and D) ERK and VEGFR2 (Y1175) phosphorylation after knockdown of the indicated phosphatases shown by immunoblotting (C). Quantification of pERK activation is shown in (D) (n = 3, mean ± SD, *p < 0.05). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23639442), licensed under a CC-BY license. Not internally tested by R&D Systems.