Human EPCR Antibody

Detection of Human EPCR by Western Blot.

Western blot shows lysates of human skin tissue, human placenta tissue, and HUVEC human umbilical vein endothelial cells. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human EPCR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2245) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for EPCR at approximately 40-45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

EPCR in Human Liver.

EPCR was detected in immersion fixed paraffin-embedded sections of human liver using 1.7 µg/mL Goat Anti-Human EPCR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2245) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of EPCR by Immunocytochemistry/ Immunofluorescence

Effect of hypoxia on endothelial hemoglobin & EPCR expression.(A) Representative WB of human cardiac microvascular endothelial cultured under hypoxia (1% oxygen) showing HIF1A, NRF2, EPCR, HBA, & ACTB as a protein loading control; n = 3 biological replicates. Values under each blot image indicate fold changes versus results in control (as determined by normalization to the loading control run on same blot). (B & C) Representative immunofluorescent images & quantification of human microvascular endothelial cultured under hypoxia at different time points, as indicated, & stained for HBA (red) & NRF2 (green). Small image in 24-hour NRF2 panel shows overlay image of the cell. *P < 0.05 & ****P < 0.0001 versus control calculated using 1-way ANOVA, Bonferroni’s multiple-comparison test. (D) Representative immunofluorescent high-resolution images showing human microvascular endothelial cultured under hypoxia (1% oxygen) & stained for PAR1 (green) & EPCR (purple) at the left panel or for PAR1 (green), EPCR (purple), & lysosomal tracker (Lyso; red) at the right panel; n = 3 biological replicates. Scale bars: 10 μm. (E & F) WB & quantification analysis of human microvascular endothelial cultured under hypoxia (1% oxygen) showing NRF2, caveolin-1, & beta-actin (ACTB) as a loading control; n = 3 biological replicates. ***P < 0.001 & ****P < 0.0001; 1-way ANOVA, Bonferroni’s multiple-comparison test. (G & H) Representative immunofluorescent images & quantification analysis showing human microvascular endothelial cultured under hypoxia (1% oxygen), treated with EPCR-blocking antibody (20 μg/mL), as indicated, & stained for HBA (red) & EPCR (purple). Scale bars: 10 μm. **P < 0.01, ***P < 0.001, & ****P < 0.0001 versus control ; 1-way ANOVA, Bonferroni’s multiple-comparison test. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35700057), licensed under a CC-BY license. Not internally tested by R&D Systems.