Human E-Selectin/CD62E Antibody

E-Selectin/CD62E in Human Kidney Cancer Tissue.

E-Selectin/CD62E was detected in immersion fixed paraffin-embedded sections of human kidney cancer tissue using Goat Anti-Human E-Selectin/ CD62E Polyclonal Antibody (Catalog # BBA18) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained (brown) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

E-Selectin/CD62E in LPS treated Human Kidney Tissue.

E-Selectin/CD62E was detected in acetone fixed cryosections of human kidney tissue using Goat Anti-Human E-Selectin/CD62E Polyclonal Antibody (Catalog # BBA18) overnight at 4 ° C. Tissues were stained with rabbit anti-goat secondary antibody and HRP polymer-conjugated anti-rabbit IgG followed by AEC+Substrate Chromogen (red) followed by counterstaining with hematoxylin (blue). Before incubation with the primary antibody, tissues were treatedex vivowith LPS and also treated to block endogenous peroxidase activity. The lower panel shows an absence of labeling when primary antibody is omitted. Experiments were carried out and images were provided by Dr. Grietje Molema, University of Groningen, The Netherlands.

Detection of E-Selectin/CD62E by Immunohistochemistry

Neutrophil transmigration hotspots are located at ICAM‐1 high expressing cellsSimplified workflow for photoconversion experiments. (1) Neutrophil flow assays were performed in widefield mode with overnight TNF alpha‐treated HUVECs expressing mEos4b. (2) The same field of views were imaged in confocal mode. (3) Areas where hotspots had appeared in the widefield video were converted from green to red using a 405 nm laser. (4) The slides were fixed and the nuclei and an adhesion molecule were stained. The original fields of view were found backing by looking for red signal. (5) In the original video, all successful and unsuccessful adhesion events were annotated. (6) The adhesion spots were overlayed with the stained image. Scale bar, 100 μm.Inverted greyscale LUT of immunofluorescence stains on overnight TNF alpha‐treated HUVECs for ICAM‐1, ICAM‐2, VCAM‐1, and E‐selectin of areas of which time lapses were made, relocated by photoconversion of mEos4b. With yellow asterisks, adhesion events that led to diapedesis are shown. With magenta asterisks, adhesion events that were followed by detachment are shown. Scale bar, 100 μm.Quantification of adhesion events on overnight TNF alpha‐treated HUVEC cells ranked by their preference for diapedesis for expression levels of ICAM‐1 (green dots, 254 diapedesis adhesion events and 57 detachment adhesion events), ICAM‐2 (magenta dots 129 diapedesis adhesion events and 79 detachment adhesion events) VCAM‐1 (blue dots 163 diapedesis adhesion events and 72 detachment adhesion events) and E‐selectin (dark yellow dots = 199 diapedesis adhesion events and 78 detachment adhesion events). Data shown are from 9 (ICAM‐1 and VCAM‐1) or 6 (ICAM‐2 and E‐selectin) images from 3 biological replicates. Median with 95% CI is shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36382783), licensed under a CC-BY license. Not internally tested by R&D Systems.