In a functional ELISA, 1-3 µg/mL of this antibody will block 50% of the binding of 50 ng/mL of Recombinant Human Angiopoietin-4 (Catalog # 964-AN) to immobilized Recombinant Human Tie-2 Fc Chimera (Catalog # 313-TI) coated at 4 µg/mL (100 µL/well). At 50 μg/mL, this antibody will block >85% of the binding.
|Angiopoietin‑4 in Human Lung. Angiopoietin‑4 was detected in immersion fixed paraffin-embedded sections of human lung using 10 µg/mL Goat Anti-Human Angiopoietin‑4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF964) 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 with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
GITR (glucocorticoid-induced TNF receptor superfamily-related protein, also named AITR, activation-inducible TNF receptor superfamily-related protein) and GITR ligand (GITRL) are novel members of the TNF receptor (TNFR) and TNF superfamilies (SF) that have been designated TNFRSF18 and TNFSF18, respectively. Human GITRL cDNA encodes a 177 amino acid residues type II membrane protein. The carboxy-terminal extracellular domain shows sequence identity to TNF/TNFSF2 (21%), Fas ligand/TNFSF6 (21%), TRAIL/TNFSF10 (18%), and lymphotoxin alpha /TNFSF1 (18%). GITRL is constitutively expressed in human umbilical vein endothelial cells but is not expressed in resting or stimulated T cell lines, B cell lines or peripheral blood mononuclear cells. GITR, the receptor for GITRL, is expressed at low levels in peripheral blood T cells, bone marrow, thymus, spleen and lymph nodes. In contrast to mouse GITR, expression of human GITR is not induced by treatment with dexamethasone, but is up-regulated by antigen-receptor stimulation or by treatment with soluble anti-CD3 plus anti-CD28 or PMA plus ionomycin. Ligation of GITR has been found to induce nuclear factor (NF)-kappa B activation via TNF receptor-associated factor 2 and protect cells from TCR activation-induced cell death. It has been proposed that GITRL and GITR may modulate T lymphocyte functions in peripheral tissues.
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