Confocal immunofluorescent analysis of stomach (left) and liver (right) using Ghrelin (D4K1W) Rabbit mAb (green). Actin filaments were labeled with DyLight 554™ Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Recommended Fluorochrome-conjugated Anti-Rabbit secondary antibodies:
NOTE: When using any primary or fluorochrome-conjugated secondary antibody for the first time, titrate the antibody to determine which dilution allows for the strongest specific signal with the least background for your sample.
Cover sections with 4% formaldehyde dilute in 1X PBS.
NOTE: Formaldehyde is toxic, use only in fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised July 2016
Protocol Id: 151
Ghrelin (D4K1W) Rabbit mAb recognizes endogenous levels of total ghrelin protein. The antibody detects acylated (n-octanoyl) and non-acylated (desacyl) forms of ghrelin (confirmed by Western blot analysis of synthetic ghrelin protein).Species Reactivity:
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of mouse ghrelin protein.
Ghrelin, also known as appetite-regulating hormone, is a neuropeptide hormone belonging to the motilin family. It is the ligand for the growth hormone secretagogue receptor type 1 (GHS-R), expressed by cells in the hypothalamic ventromedial nucleus and arcuate nucleus (1). Ghrelin is synthesized as a preprohormone by ghrelinergic cells in the gastrointestinal tract; proteolytic cleavage yields a 28-amino acid peptide hormone, which undergoes obligate n-octanoylation at serine 3 by the enzyme ghrelin O-acetyltransferase (GOAT) (2). Binding of n-octanoyl ghrelin to GHS-R stimulates growth hormone release, while simultaneously exerting multiple neuroendocrine affects that influence appetite, gastric motility and energy balance (3).
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DRAQ5 is a registered trademark of Biostatus Limited.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.