Human Endoglin/CD105 Antibody

Endoglin/CD105 in Mouse Kidney.

Endoglin/CD105 was detected in immersion fixed paraffin-embedded sections of mouse kidney using Goat Anti-Human Endoglin/CD105 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1097) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membranes in tubules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Endoglin/CD105 by Simple Western^TM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Endoglin/CD105 knockout HeLa cell line (KO), loaded at 0.2 mg/mL. A specific band was detected for Endoglin/CD105 at approximately 121 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. Goat Anti-Human Endoglin/CD105 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1097) was used at 20 µg/mL followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF019). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western Blot Shows Human Endoglin/CD105 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Endoglin/CD105 knockout HeLa cell line (KO). PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human Endoglin/CD105 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1097) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for Endoglin/CD105 at approximately 90 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Endoglin/CD105 by Immunocytochemistry/Immunofluorescence

Overview of Multi-dimensional Microscopic Molecular Profiling (MMMP).The overall MMMP approach is depicted using an example tissue section from normal human duodenum (sample #1.9.7). (a) Slides were subjected to repeated cycles of staining and imaging with fluorescent primary antibodies and DAPI. At the end of each cycle, fluorescent signal was removed by a chemical bleaching process, and slides were again imaged, before proceeding to the next round of this iterative procedure. After the final antibody stain (#15 Sma), slides were analyzed with a series of histochemical stains. (b) A set of tiling images spanning each tissue section was initially generated by the microscope system. The tiling images were then computationally ‘stitched’ together to produce a single image per staining cycle for each sample. (c) Image registration was performed to align images from the same tissue section across cycles. Mean intensities of the DAPI signal from all immuno-fluorescence images are shown from before (Unregistered) and after (Registered) the image registration procedure was completed. (d) Following registration, signal intensities from the relevant channels for each image (columns) in the MMMP series were extracted for each pixel (rows) within the tissue section and compiled into a large data matrix of in situ molecular profiles. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0128975), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Endoglin/CD105 by Flow Cytometry

Transfection of stem cells with mRNA nanoparticles can promote their expansion and self-renewal. a Targeting of CD105 enables specific transfection of HSC CD34+ cells. Cells were left untreated, or transfected with eGFP-encoding mRNA in nanoparticles (NPs) coated with PGA coupled to a control antibody or anti-CD105. Transfection efficiency was assayed by flow cytometry after 24 h. b NP transfection efficiency in CD34+ samples from three independent donors. Viability is shown in c. d Expansion of CD34 + PBSCs after NP-transfection. e Phenotypical characterization of PBSC-derived CD34+ subpopulations after 2 days in culture. Cells were either transfected with eGFP NPs on day 1 or left unmodified. Gating is indicated in brackets on top of each column. f Summary bar graph showing mean frequencies and SE of primitive Hematopoietic Stem Cells (HSCs), Multipotent Progenitors (MPPs), Lymphoid-primed Multipotent Progenitors (LMPs), and Early Myeloid Progenitors (EMPs). PBSCs from four independent donors were analyzed. Error bars represent mean ± SE. g Colony output of sort-purified GFP-NP transfected versus unmodified CD34+ cells from day 7 cultures (n = 3 cultures from independent donors); n.s., non-significant. Arising colonies were identified as colony forming unit (CFU) granulocyte (CFU-G), macrophage (CFU-M), granulocyte-macrophage (CFU-GM) and burst forming unit-erythrocyte (BFU-E). Colonies consisting of erythroid and myeloid cells were scored as CFU-MIX; n.s., non-significant. Error bars represent mean ± SE. h Representative images of CFU-MIX colonies from untransfected and GFP-NP-transfected CD34+ cells (×4-magnification; scale bar 1000 µm). i qPCR measurements of NP-delivered Musashi-2 (MSI2) mRNA expression over time. Error bars represent mean ± SE. j Comparison of CD133 and CD34 expression in HSCs transfected with control GFP mRNA-NPs versus MSI2 mRNA-NPs, assessed by flow cytometry 8 days after NP exposure and cell expansion. Data represent two independent experiments conducted in triplicate. k Cellular fold expansion of CD34− (differentiated) and CD34+ CD133+ (progenitor) cells. Bar graphs show mean and SE of three independent experiments. Data represent two independent experiments conducted in triplicate. l Colony forming unit outputs of untransfected versus MSI2-NP-transfected HSCs (n = 3 cultures from independent donors); Pairwise differences between groups were analyzed with the unpaired, two-tailed Student’s t Test. *P = 0.049, **P = 0.012, ***P = 0.011; n.s., non-significant. Error bars represent mean ± SE. m Representative images of colonies from untransfected and MSI2-NP-transfected CD34+ cells (scale bar, 300 µm) Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28855514), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Endoglin/CD105 by Immunocytochemistry/Immunofluorescence

Spatiotemporal changes in tumor vascularization and COX2 coverage at different stages of prostate cancer progression/regression. The 5-μm thick paraffin serial sections were stained with antibodies against endoglin (CD105, red), cyclooxygenase 2 (COX2, green), and CD68 (white). (A–D) Representative 200× pictures are showing changes in vascularization and COX2 expression in tertiary lymphoid organs (TLO), or (E–H) tumor areas. (A) CD105+ vessels, CD68+COX2+ macrophages, and CD68−COX2+ cells are detected in TLO from prostatic intraepithelial neoplasia (PIN) patients. (B) Smaller CD105+ vessels and CD68−COX2+ cells are located on the border of TLO from intermediate prostate cancer patient. (C) In a prostatectomy specimen from advanced carcinoma patient, CD105+ vessels with cuboidal morphology are located outside TLO, while numerous CD68−COX2+ cells are found in the center of a TLO. (D) Oval-shaped and spindle-shaped COX2+ cells are located on the border of a TLO. (E) Vascularity and COX2+ epithelial cells are modestly increased in tumor areas from PIN patients. (F,G) Abundant blood vessels with abnormal morphology and aberrant organization were located in close proximity to strongly labeled COX2+ transformed epithelium in tumors of intermediate and advanced prostate cancer patients. (D,H) Preserved vascular morphology and organization, as well as reduced COX2+ density were observed in TLO and tumor areas from patients with spontaneous prostate cancer regression. CD105+ vessels with a high endothelial venule-like morphology were detected in TLO from patients at late stages of prostate cancer (white asterisk). Representative 200× magnification pictures of TLO and tumor areas were taken with a Zeiss Axioplan Microscope and recorded with a Hamamatsu Camera. Scale bars represent 100 μm. Image collected and cropped by CiteAb from the following publication (https://journal.frontiersin.org/article/10.3389/fimmu.2017.00563/full), licensed under a CC-BY license. Not internally tested by R&D Systems.