The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/500 - 1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 21 kDa).
RelevanceGlucagon is a hormone that is secreted by alpha cells in the pancreas. Glucagon antagonizes insulin by converting glycogen to glucose in the liver and increasing blood sugar levels. Glucagon-like peptide 1 (GLP1), Glucagon-like peptide 2 (GLP2), VIP (vasoactive intestinal peptide) and PACAP (pituitary adenylate cyclase activating polypeptide) are in the glucagons hormone family. GLP1 is a transmitter in the central nervous system that regulates feeding and drinking behavior. GLP2 stimulates intestinal epithelial growth.
IHC image of Glucagon staining in human pancreas formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab53704, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Glucagon antibody (ab53704)
All lanes : Anti-Glucagon antibody (ab53704) at 1/500 dilution
Lane 1 : COS7 cell extract Lane 2 : COS7 cell extract with immunising peptide