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. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration.
1/100. Perform heat mediated antigen retrieval via the pressure cooker method (1 minute, with 1 mmol EDTA at pH8) before commencing with IHC staining protocol.
Use a concentration of 0.4 µg/ml. (0.4 µg/well)
Plays a major role in ketone body metabolism.
Involvement in disease
Defects in ACAT1 are a cause of 3-ketothiolase deficiency (3KTD) [MIM:203750]; also known as alpha-methylacetoaceticaciduria. 3KTD is an inborn error of isoleucine catabolism characterized by intermittent ketoacidotic attacks associated with unconsciousness. Some patients die during an attack or are mentally retarded. Urinary excretion of 2-methyl-3-hydroxybutyric acid, 2-methylacetoacetic acid, triglylglycine, butanone is increased. It seems likely that the severity of this disease correlates better with the environmental or acquired factors than with the ACAT1 genotype.
Immunocytochemistry image of ab110290 stained human HDFn cells. The cells were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15 min). The cells were incubated with the antibody (9H10AB4, 5 µg/ml) for 2 hours at room temperature or over night at 4°C. The secondary antibody was (red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1 hour. 10% Goat serum was used as the blocking agent for all blocking steps. DAPI was used to stain the cell nuclei (blue). Target protein locates mainly in mitochondria.
ACAT1 immunohistochemistry in human cerebellum visualized with ab110290. ACAT1 immunoactivity is most intense in neuronal cell bodies, most notably in the large Purkinje cells. Note the distinctive subcellular localization of ACAT1 immunoreactivity in the Purkinje cell bodies. The functional significance of this pattern is unknown at present but this antibody offers the opportunity to investigate it in more detail.