The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. Preliminary experiments in Human Pancreas lysates gave no specific signal but low background (at antibody concentration up to 1µg/ml).
Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use at an assay dependent concentration. Antibody detection limit for peptide ELISA 1:4000
Use a concentration of 5 µg/ml.
The cholecystokinin (CCK) family of peptide hormones have been implicated in numerous important physiologic events. These appear to be mediated through 2 general classes of receptors, A (CCKAR)and B (CCKBR), based on their binding affinities for CCK/gastrin family peptides. Through binding to class A receptors, CCK is a major physiologic mediator of gallbladder contraction and pancreatic enzyme secretion. It appears to play a role in slowing gastric emptying, relaxation of the sphincter of Oddi, and potentiation of insulin secretion. Further, it has been implicated as a mediator of pancreatic growth and tumorigenesis. Class A receptors have also been described in the anterior pituitary, myenteric plexus, and regions of the central nervous system, where they have been implicated in the pathogenesis of feeding disorders, Parkinson disease, schizophrenia, and drug addiction.
ICC/IF image of ab77269 stained CaCo2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab77269 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 donkey anti- goat (ab96931) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
El-Kouhen K & Morisset J Control of somatostatin (SS) secretion by CCK-1 and CCK-2 receptors' occupation in RIN-14B cells, a rat pancreatic islet cell line. Pancreas39:127-34 (2010).
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