Monoclonal antibody 2D7 recognizes the pro-region of Urocortin-2. Urocortins 1, 2 and 3 (Ucn I, Ucn II, and Ucn III), members of the corticotropin releasing factor (CRF)4 family, have diverse bioactivities and participate in the neuroendocrine responses to stress, appetite suppression, immunomodulation, energy and water homeostasis, and cardiovascular function. Their actions are mediated through the 7-transmembrane G-protein coupled receptors CRFR1 and CRFR2. Ucn1 binds to both receptors, whereas Ucn II and Ucn III are specific to CRFR2, widely expressed in the brain, heart, vasculature, and peripheral tissues. Binding of Ucn II to its receptor triggers an intricate signaling cascade via protein kinase C, phosphatidylinositol-3-kinase/Akt and mitogen-activated protein kinase pathways to modulate cytokine expression, cell proliferation, and apoptotic responses. The presence of Ucn II and CRFR2 in the heart and vasculature suggest a potentially important role for Ucn II in the physiology and pathophysiology of the circulation in health and cardiovascular disease. Urocortins exert beneficial effects in experimental cardiovascular disease models and may have therapeutic potential. The sequence of the mature human Ucn II neuropeptide is IVLSLDVPIGLLQILLEQARARAAREQATTNARILARV.
Immuno assays, Western blot
IA: Antibody 2D7 can be used as capture antibody (Ref.1).
W: Protein samples were separated by 16% SDS-PAGE for an hour at 200 V and electroblotted onto Immun-Blot® PVDF membrane at 100 mA for 1 h. Western blotting was performed using mouse 2D7 (1/1000 dilution) as primary antibody and goat anti mouse IgG conjugated to horse radish peroxidase (1/2000). The band sizes are 35 and 80 kDa. (Ref.1)
Dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.