polyclonal goat anti epcr antibodies  (R&D Systems)

Journal: PLoS ONE

Article Title: Blood outgrowth endothelial cells (BOECs) as a novel tool for studying adhesion of Plasmodium falciparum -infected erythrocytes

doi: 10.1371/journal.pone.0204177

Figure Lengend Snippet: (A) Process of isolating BOECs from human blood samples. B) Bright field images showing 48 hours post-seeding and documenting the appearance of progenitors (black arrow, day 7), small colonies (white arrow, day 14) and confluent BOEC (day 28) over time. C) Receptor expression (CD201/EPCR; PECAM-1/CD31; thrombomodulin/CD141 and CD54/ICAM-1) on resting and activated BOECs at passage 3. Shown is a representative immunofluorescence image (400 times magnification at passage 3) from a single donor. Bright field image (200 times magnification) shows elongation of cells after TNFα treatment.

Article Snippet: Mouse monoclonal antibodies against human MUC18 (CD146; clone P1H12, Biolegend), PECAM-1 (CD31; clone 9G11, R&D systems), ICAM-1 (CD54; clone BBIG-11, R&D systems), thrombomodulin (CD141; clone 501733, R&D systems), platelet glycoprotein 4 (CD36; clone FA6.152, Beckman Coulter), and goat polyclonal antibody against human EPCR (CD201; polyclonal R&D systems) were used as primary antibodies.

Techniques: Expressing, Immunofluorescence

Journal: PLoS ONE

Article Title: Blood outgrowth endothelial cells (BOECs) as a novel tool for studying adhesion of Plasmodium falciparum -infected erythrocytes

doi: 10.1371/journal.pone.0204177

Figure Lengend Snippet: Surface expression of ICAM-1/CD54, EPCR/CD201, CD36, PECAM-1/CD31, CD141, MUC18/CD146 and prominin-1/CD133 on resting (grey histograms, left column) or TNFα-activated BOECs (grey histograms, middle column) and secondary only (black histograms, left and middle column) from one representative Ghanaian child (P360; passage 3–6). Right column: mean fluorescence intensity (MFI) of eight individual BOEC cells lines (passage 3–6) either resting or TNFα-activated. Data from three independent experiments performed in duplicate.

Article Snippet: Mouse monoclonal antibodies against human MUC18 (CD146; clone P1H12, Biolegend), PECAM-1 (CD31; clone 9G11, R&D systems), ICAM-1 (CD54; clone BBIG-11, R&D systems), thrombomodulin (CD141; clone 501733, R&D systems), platelet glycoprotein 4 (CD36; clone FA6.152, Beckman Coulter), and goat polyclonal antibody against human EPCR (CD201; polyclonal R&D systems) were used as primary antibodies.

Techniques: Expressing, Fluorescence

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Renal endothelial protein C receptor expression and shedding during diabetic nephropathy.

doi: 10.1111/jth.13315

Figure Lengend Snippet: Fig. 1. Plasma and urinary soluble endothelial protein C receptor (sEPCR) concentrations and Spearman correlation analysis. Plasma and uri- nary sEPCR levels were measured by ELISA. (A–C) Plasma (A) and urine (B) sEPCR levels and 24-h urinary excretion (C) are elevated in patients suffering from diabetic nephropathy (DN) as compared with diabetic controls. (D) Twenty-four-hour urinary sEPCR excretion corre- lates with plasma sEPCR level in the whole population. (E, F) Urinary sEPCR excretion does not correlate with glomerular filtration rate (GFR) in the diabetic control group (E) but does in the DN group (F). *P < 0.05 (Mann–Whitney test). **P < 0.01 (Mann–Whitney test).

Article Snippet: The membranes were blocked for 1 h in 20 mM Tris, 150 mM NaCl, and 1% Tween, pH 7.5/5% non-fat dry milk, and incubated overnight at 4 °C with the primary antibodies, including goat anti-human EPCR polyclonal antibody (R&D Systems) and rabbit anti-human ADAM-17 polyclonal antibody (LSBio).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, MANN-WHITNEY

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Renal endothelial protein C receptor expression and shedding during diabetic nephropathy.

doi: 10.1111/jth.13315

Figure Lengend Snippet: Fig. 2. Endothelial protein C receptor (EPCR) expression in renal biopsies. (A–D) Representative EPCR immunostaining of kidney biopsies (A, B), with close-ups of glomeruli (C, D). (E–G) Diabetic nephropathy (DN) patients show markedly reduced EPCR expression on glomerular endothelium (E), whereas ECPR staining intensity remained unchanged in podocytes (F) and mesangium (G) as compared with healthy con- trols. (H) Tubular ECPR staining intensity did not differ between the two groups. In addition, an internal control (extraglomerular vessels) remained strongly stained in DN tissue, confirming that the staining was technically successful. Results are shown as mean and standard error of the mean. *P < 0.05 (Mann–Whitney test).

Article Snippet: The membranes were blocked for 1 h in 20 mM Tris, 150 mM NaCl, and 1% Tween, pH 7.5/5% non-fat dry milk, and incubated overnight at 4 °C with the primary antibodies, including goat anti-human EPCR polyclonal antibody (R&D Systems) and rabbit anti-human ADAM-17 polyclonal antibody (LSBio).

Techniques: Expressing, Immunostaining, Staining, Control, MANN-WHITNEY

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Renal endothelial protein C receptor expression and shedding during diabetic nephropathy.

doi: 10.1111/jth.13315

Figure Lengend Snippet: Fig. 3. Endothelial protein C receptor (EPCR) shedding in glomerular endothelial cells (GEnCi). (A, B) Elevated EPCR shedding was induced when GEnCi cells were treated with 1 lM phorbol 12-myristate 13-acetate (PMA) (A) or incubated in high-glucose (30 mM) medium (B). (C) ADAM-17 was inhibited by 50 lM TAPI-0 (B) or ADAM-17 small interfering RNA (siRNA). (D) Representative western blot analysis for EPCR and ADAM-17. (E, F) Quantification of protein expression of ADAM-17 (E) and EPCR (F) in GEnCi cells incubated under control conditions (Vehicle), with the osmotic control mannitol, in high-glucose medium and following ADAM-17 inhibition by TAPI-0. (G–L) mRNA expression of vascular endothelial cadherin (VE-cadherin), a marker of endothelial phenotype, and transforming growth factor (TGF)-b, a mar- ker of endothelial phenotype loss, in GEnCi cells incubated under control conditions (Vehicle), with the osmotic control mannitol, in high-glu- cose medium and following ADAM-17 inhibition by TAPI-0 (G, H), ADAM-17 siRNA (I, J) and EPCR siRNA (K, L). Results are shown as mean and standard error of the mean. *P < 0.05 (Mann–Whitney test). GFP, green fluorescent protein; sEPCR, soluble EPCR.

Article Snippet: The membranes were blocked for 1 h in 20 mM Tris, 150 mM NaCl, and 1% Tween, pH 7.5/5% non-fat dry milk, and incubated overnight at 4 °C with the primary antibodies, including goat anti-human EPCR polyclonal antibody (R&D Systems) and rabbit anti-human ADAM-17 polyclonal antibody (LSBio).

Techniques: Incubation, Small Interfering RNA, Western Blot, Expressing, Control, Inhibition, Marker, MANN-WHITNEY

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Renal endothelial protein C receptor expression and shedding during diabetic nephropathy.

doi: 10.1111/jth.13315

Figure Lengend Snippet: Fig. 4. ADAM-17 expression in renal tissue. (A–D) Representative immunostaining of kidney biopsies for ADAM-17 in glomeruli (A, B), with close-ups of a glomerular segment (C, D). Podocytes are marked with an arrow, endothelium with a V, and mesangium with #. (E) Diabetic nephropathy (DN) patients have elevated glomerular endothelium expression of ADAM-17 as compared with healthy controls. (F, G) EPCR expression was similar on podocytes and mesangium in the two groups. Results are shown as mean and standard error of the mean. *P < 0.05 (Mann–Whitney test).

Article Snippet: The membranes were blocked for 1 h in 20 mM Tris, 150 mM NaCl, and 1% Tween, pH 7.5/5% non-fat dry milk, and incubated overnight at 4 °C with the primary antibodies, including goat anti-human EPCR polyclonal antibody (R&D Systems) and rabbit anti-human ADAM-17 polyclonal antibody (LSBio).

Techniques: Expressing, Immunostaining, MANN-WHITNEY