Immunofluorescence was performed on sections of adult rat pineal. Tissue sections on slides were probed for 24 h at 4°C in a humidified chamber with rabbit anti-AANAT (Product # PA5-26631) at an antibody concentration of 1-2 µg/ml diluted in 50 mM Tris-HCl, pH 7.4, containing 1.5% NaCl, 0.3% Triton X-100 (TBST) and 4% normal goat serum. After overnight incubation, sections were washed extensively for 1 h in TBST, and detection of primary antibody was performed for 1.5 h at room temperature with AlexaFluor-488-conjugated donkey anti-rabbit diluted 1:600 in TBST. Sections were then washed in TBST, then in TBS (without triton) and then coversliped with anti-fade medium. Images were taken on a Zeiss Z2 scanning microscope at x40 objective magnification, and show immunofluorescence labelling of AANAT localized to pinealocytes. Data courtesy of Dr. James Nagy's laboratory.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||KLH conjugated synthetic peptide between 27-54 amino acids from the N-terminal region of human AANAT|
|Purification||Antigen affinity chromatography|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1-2 µg/ml|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA5-26631 has successfully been used to detect serotonin N-AT in both WB and IF applications.
AANAT belongs to the acetyltransferase superfamily. It is the penultimate enzyme in melatonin synthesis and controls the night/day rhythm in melatonin production in the vertebrate pineal gland. Melatonin is essential for the function of the circadian clock that influences activity and sleep. This enzyme is regulated by cAMP-dependent phosphorylation that promotes its interaction with 14-3-3 proteins and thus protects the enzyme against proteasomal degradation. This gene may contribute to numerous genetic diseases such as delayed sleep phase syndrome.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.