The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 28 kDa (predicted molecular weight: 30 kDa).
Enkephalin and enkephalin-like peptides have been detected in many animal species. In mammals these opioid pentapeptides are cleaved from a common precursor molecule which is expressed in a number of cell types, including neurons and endocrine cells associated with the sympathetic nervous system where they are often co-stored with catecholamines. They are known to be endogenous ligands for Sopioid receptors and are known to mediate analgesia at the spinal level.
Immunocytochemistry/ Immunofluorescence - Leu Enkephalin antibody (ab85798)
ICC/IF image of ab85978 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat 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 (ab85798, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.