Human LYVE-1 Antibody

LYVE‑1 in Human Tonsil. LYVE‑1 was detected in immersion fixed paraffin-embedded sections of human tonsil using 15 µg/mL Goat Anti-Human LYVE‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2089) 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 Human LYVE‑1 by Western Blot. Western blot shows lysates of human liver and spleen tissue. PVDF membrane was probed with 0.25-1 µg/mL of Goat Anti-Human LYVE-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2089) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for LYVE-1 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of Human LYVE-1 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 LYVE-1 by Western Blot TGF-beta 1, -beta 2 and -beta 3 reduce lymphatic marker expression in LECs.Primary human LECs were treated with 10, 20 or 30 ng/ml TGF-beta 1, -beta 2 and -beta 3 for 72 hours (a) or 100 hours (b). Untreated cells served as a control. Lysates were prepared and analysed by Western blot using antibodies specific for Lyve-1, Prox-1, VEGFR-3 or vimentin. Vinculin served as loading control. The experiment was performed twice with equivalent results. For densitometry evaluation, protein bands were analysed using the software ImageJ. Bands for the Prox-1, Lyve-1, vimentin and VEGFR-3 proteins were normalized to the corresponding loading control and are displayed as the expression level relative to the untreated control samples. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0162221), licensed under a CC-BY license. Not internally tested by R&D Systems.