Mouse/Rat DLL1 Antibody

Detection of DLL1 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Sheep Anti-Mouse/Rat DLL1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3970, filled histogram) or control antibody (5-001-A, open histogram), followed by NorthernLights™ 637-conjugated Anti-Sheep IgG Secondary Antibody (NL011).

DLL1 in Embryonic Mouse Stomach. DLL1 was detected in immersion fixed frozen sections of embryonic mouse stomach (E13.5) using 10 µg/mL Sheep Anti-Mouse/Rat DLL1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3970) overnight at 4 °C. Tissue was stained with the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Detection of Mouse DLL1 by Immunocytochemistry/Immunofluorescence Mpdz does not affect cell cell junction assembly.(A, B) HUVECs were either transduced with lentivirus expressing GFP (sh-ctrl) or with lentivirus expressing shRNA against MPDZ (sh-MPDZ). Cells were cultured under sparse conditions (A) or confluent conditions (B). After PFA fixation cells were stained for DLL1 and Nectin-2 or DLL4 and Nectin-2 and counterstained with DAPI. Images were acquired with the confocal microscope LSM 700. Arrow indicates co-localization of DLL1/4 with Nectin-2 at the cell membrane. Arrow head indicates diminished co-localization at the cell membrane. Scale bar: 10 µm. (C) HUVECs were either transfected with control siRNA (si-ctrl) or with siRNA against Nectin-2 (si-Nectin-2). After PFA fixation cells were stained for DLL1 and Nectin-2 or DLL4 and Nectin-2. Images were acquired with the confocal microscope LSM 700. Arrow indicates localization of DLL1/4 at the cell membrane.Scale bar: 10 µm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29620522), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse DLL1 by Western Blot MPDZ promotes Notch signaling activity.(A) HUVECs were either transduced with lentivirus expressing GFP (sh-ctrl) or with lentivirus expressing shRNA against MPDZ (sh-MPDZ). Expression level of Notch target genes HEY1, HEY2 and HES1 were analyzed by qPCR 48 hr after transduction. Data are presented as mean ±SD. n ≥ 3; *, p<0.05; **, p<0.01; ***, p<0.001 unpaired Student’s t-test. (B) Cardiac endothelial cells were isolated from Mpdzfl/fl and Mpdz delta EC mice by magnetic beads bound with CD31 antibodies. Expression levels of Notch target genes Hey1 and Hey2 were analyzed by qPCR. Data are presented as mean ±SD. n = 3; *, p<0.05; ***, p<0.001 unpaired Student’s t-test. (C) HUVECs were either transduced with lentivirus expressing GFP (sh-ctrl) or with lentivirus expressing shRNA against MPDZ (sh-MPDZ). Expression levels of DLL1 and DLL4 were analyzed by immunoblotting 48 hr after transduction. beta -actin served as loading control. Data are presented as mean ±SD. n ≥ 3; n.s., not significant. (D) HUVECs were either transduced with adenovirus expressing GFP (ctrl) or with adenovirus expressing MPDZ. Expression levels of DLL1 and DLL4 were analyzed by immunoblotting 48 hr after transduction. beta -actin served as loading control. Data are presented as mean ±SD. n ≥ 3; n.s., not significant. (E) Lung endothelial cells were isolated from Mpdzfl/fl and Mpdz delta EC mice by CD31 magnetic beads. Protein amounts of Dll1 and Dll4 were analyzed by immunoblotting. beta -actin served as loading control. Data are presented as mean ±SD. n = 3; n.s., not significant.10.7554/eLife.32860.007Figure 2—source data 1.Source data of qantitative PCR analysis related to Figure 2A and B.Source data of qantitative PCR analysis related to Figure 2A and B. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29620522), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse DLL1 by Western Blot MPDZ promotes Notch signaling activity.(A) HUVECs were either transduced with lentivirus expressing GFP (sh-ctrl) or with lentivirus expressing shRNA against MPDZ (sh-MPDZ). Expression level of Notch target genes HEY1, HEY2 and HES1 were analyzed by qPCR 48 hr after transduction. Data are presented as mean ±SD. n ≥ 3; *, p<0.05; **, p<0.01; ***, p<0.001 unpaired Student’s t-test. (B) Cardiac endothelial cells were isolated from Mpdzfl/fl and Mpdz delta EC mice by magnetic beads bound with CD31 antibodies. Expression levels of Notch target genes Hey1 and Hey2 were analyzed by qPCR. Data are presented as mean ±SD. n = 3; *, p<0.05; ***, p<0.001 unpaired Student’s t-test. (C) HUVECs were either transduced with lentivirus expressing GFP (sh-ctrl) or with lentivirus expressing shRNA against MPDZ (sh-MPDZ). Expression levels of DLL1 and DLL4 were analyzed by immunoblotting 48 hr after transduction. beta -actin served as loading control. Data are presented as mean ±SD. n ≥ 3; n.s., not significant. (D) HUVECs were either transduced with adenovirus expressing GFP (ctrl) or with adenovirus expressing MPDZ. Expression levels of DLL1 and DLL4 were analyzed by immunoblotting 48 hr after transduction. beta -actin served as loading control. Data are presented as mean ±SD. n ≥ 3; n.s., not significant. (E) Lung endothelial cells were isolated from Mpdzfl/fl and Mpdz delta EC mice by CD31 magnetic beads. Protein amounts of Dll1 and Dll4 were analyzed by immunoblotting. beta -actin served as loading control. Data are presented as mean ±SD. n = 3; n.s., not significant.10.7554/eLife.32860.007Figure 2—source data 1.Source data of qantitative PCR analysis related to Figure 2A and B.Source data of qantitative PCR analysis related to Figure 2A and B. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29620522), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of DLL1 in transfected CHO cells by Flow Cytometry CHO cells transfected with DLL1 were stained with Sheep Anti-Mouse/Rat DLL1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3970, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram) followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127). View our protocol for Staining Membrane-associated Proteins.

Detection of DLL1 in Embryonic Mouse Stomach (E13). DLL1 was detected in perfusion fixed frozen sections of Embryonic Mouse Stomach (E13) using Sheep Anti-Mouse/Rat DLL1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3970) at 1 µg/ml overnight at 4 °C. Tissue was stained using the HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.