The antibody produces strong labeling of GHRH at a dilution of 1/2,000 – 1/4,000 using biotin/streptavidin HRP in rat hypothalamus (median eminence). Optimal dilution will vary depending upon fixation, labeling tehcnique and/or detection system; therefore, a dilution series is recommended.
Cross reactivity of GHRF antiserum was examined using the paper spot technique of Larsson (1981). Using 2 µL, 100 pmole amounts, the following substances did not react with rat GHRF antisera diluted 1/500 using the PAP labeling method: glucagon, gastric inhibitory peptide, secretin, vasoactive intestinal peptide, peptide histidine isoleucine, pancreatic polypeptide (human or rat), human GHRF, somatostatin, insulin, ACTH, motilin, cholecystokinin octapeptide, substance P, molluscan cardioexcitatiory peptide, gastrin 34, and serotonin. GHRF antiserum had a very good reactivity using rat GHRF at 2 µL, 100 pmole amounts.
Photo Description: IHC image of neurons staining for DBH in the rat brainstem. The tissue was fixed with 4% formaldehyde/0.05% glutaraldehyde in 0.1 M phosphate buffer, before being removed and prepared for vibratome sectioning. Floating sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. Primary antibody was incubated at 1:4000 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen.
Quantity / Volume: 100 µL
Reacts With: Rat
Alternate Names: GRF; GHRH; Somatoliberin; Growth hormone-releasing hormone; Sermorelin; Somatocrinin; Somatorelin; Ghrf; growth hormone releasing hormone, anti-GHRF
Gene Symbol: Ghrh
Entrez Gene: 29446 Rat