mGLUR4 [UMB18] (RM-non-phospho-Metabotropic Glutamate Receptor 4 Antibody), Rabbit Monoclonal

Figure 1. Indentification of the Endothelin  Receptor A in mouse tissues. Native mouse tissues were lysed and GPCR receptors were enriched by using WGA-beads (see 7TM Western Blot Protocol). Extracted proteins were immunoblotted with the phosphorylation-independent anti-ETA antibody (7TM00219N-RM) at a dilution of 1:1000.

Figure 2. Immunohistochemical identification of Endothelin Receptor A in mouse heart. Sections were dewaxed, microwaved in citric acid, and incubated with anti-ETA (Endothelin receptor A) antibody (7TM0219N-RM) at a dilution of 1:100. Sections were then sequentially treated with biotinylated anti-rabbit IgG and avidin-biotin solution.Color was developed by incubation in 3-amino-9-ethylcarbazole (AEC), and sections were counterstained with hematoxylin. Note, ETA receptors were detected at the plasma membrane of cardiomyocytes.

Figure 3. Immunohistochemical identification of Endothelin Receptor A in human ovarian carcinoma. Sections were dewaxed, microwaved in citric acid, and incubated with anti-ETA (Endothelin receptor A) antibody (7TM0219N-RM) at a dilution of 1:100. Sections were then sequentially treated with biotinylated anti-rabbit IgG and avidin-biotin solution. Sections were then developed in 3,3-diaminobenzidine (DAB)-glucose oxidase and lightly counterstained with hematoxylin. Note, ETA receptors were uniformly detected at the plasma membrane of tumor cells.

Figure 4. Immunocytochemical identification of Endothelin Receptor A in HEK293 cells. HEK293 cells stably expressing the Endothelin Receptor A (ETA) were either not exposed or exposed to 1 μM Endothelin-1 for 30 min and immunocytochemically stained with anti-ETA (Endothelin Receptor A) antibody (7TM0219N-RM) at a dilution of 1:200. Note, ETA receptors were confined to the plasma membrane in untreated cells (0 min). ETA receptors were seen in perinuclear clusters of vesicles after 30 min Endotheline-1 exposure.

Figure 5. Validation of the Endothelin Receptor A in transfected HEK293 cells. Native HEK293 cells (MOCK) or HEK293 cells stably expressing the Endotheline Receptor A (ETA) were lysed and immunoblotted with the phosphorylation-independent anti-ETA antibody (7TM0219N-RM) at a dilution of 1:1000.

Lupp A, Mann A, Heeb A, Kaemmerer D, Sänger J, Evert M, Evert K, Mawrin C, Schulz S. Reassessment of endothelin receptor A expression in normal and neoplastic human tissues using the novel rabbit monoclonal antibody UMB-8. Peptides. 2015 Apr;66:19-25. doi: 10.1016/j.peptides.2015.02.005. Epub 2015 Feb 20. PubMed PMID: 25708416.