Overview

  • Product name

  • Description

    Rabbit polyclonal to AANAT
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Rat
    Predicted to work with: Mouse, Chicken, Hamster, Human, Chimpanzee, Rhesus monkey
  • Immunogen

    Synthetic peptide corresponding to Mouse AANAT aa 24-33 (N terminal).
    Sequence:

    CQRRHTLPAS


    (Peptide available as ab5003)

  • Positive control

    • Rat pineal gland whole protein extract IHC-P: FFPE Rat brain tissue sections.
  • General notes

    .

     This product was previously labelled as Serotonin N-acetyltransferase

     

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Total anti-N-terminal AA-NAT-specific IgG was affinity purified on resin-coupled non-phosphopeptide. Phospho-specific amino-terminal IgG was subsequently isolated on resin-coupled phosphopeptide.
  • Primary antibody notes

    Pineal gland serotonin N-acetyltransferase (arylalkylamine N-acetyltransferase, AA-NAT) is an ~23 kDa protein which is the rate-limiting enzyme in melatonin synthesis. Melatonin serves as the hormonal signal of the daily light/dark cycle, or body clock. The nocturnal oscillation in enzyme activity ranges by 10- to 100-fold depending on species among vertebrates. Studies have shown that AA-NAT is regulated by cAMP at both the mRNA and protein levels. AA-NAT has putative protein kinase A (PKA) sites at Thr 29 and Ser 206. Phosphorylation of AA-NAT at these residues might play a role in protein inactivation via proteasome-dependent degradation.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab3505 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 0.5 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).Can be blocked with AANAT peptide (ab5003).

This antibody detects an ~23 kDa protein representing unphosphorylated and phosphorylated AA-NAT from rat pineal gland whole protein extract.

IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    Controls the night/day rhythm of melatonin production in the pineal gland. Catalyzes the N-acetylation of serotonin into N-acetylserotonin, the penultimate step in the synthesis of melatonin.
  • Tissue specificity

    Highly expressed in pineal gland and at lower levels in the retina. Weak expression in several brain regions and in the pituitary gland.
  • Pathway

    Aromatic compound metabolism; melatonin biosynthesis; melatonin from serotonin: step 1/2.
  • Involvement in disease

    Delayed sleep phase syndrome
  • Sequence similarities

    Belongs to the acetyltransferase family. AANAT subfamily.
    Contains 1 N-acetyltransferase domain.
  • Post-translational
    modifications

    cAMP-dependent phosphorylation on both N-terminal Thr-31 and C-terminal Ser-205 regulates AANAT activity by promoting interaction with 14-3-3 proteins.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Database links

  • Alternative names

    • AA-NAT antibody
    • Aanat antibody
    • Aralkylamine N-acetyltransferase antibody
    • Arylalkylamine N acetyltransferase antibody
    • Serotonin acetylase antibody
    • Serotonin N-acetyltransferase antibody
    • SNAT antibody
    • SNAT_HUMAN antibody
    see all

Images

  • IHC image of AANAT staining in Rat brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab3505, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:

  • Chen L  et al. Pinealectomy or light exposure exacerbates biliary damage and liver fibrosis in cholestatic rats through decreased melatonin synthesis. Biochim Biophys Acta Mol Basis Dis 1865:1525-1539 (2019). Read more (PubMed: 30890428) »
  • Hou B  et al. Homeostatic Plasticity Mediated by Rod-Cone Gap Junction Coupling in Retinal Degenerative Dystrophic RCS Rats. Front Cell Neurosci 11:98 (2017). WB, IHC (PFA fixed) ; Rat . Read more (PubMed: 28473754) »
See all 3 Publications for this product

Customer reviews and Q&As

Answer

Regarding the feedback on the secondary and the loading control working well, this is very reassuring and tells us the problem is not due to these. As the customer is sure the cells she is investigating are still pineal and should definitely express Serotonin N-acetyltransferase we would indeed expect a band at the correct size in her samples. I have read through Tanya's recommendations and agree with her that blocking can make a big difference. In our laboratory we try various blocking agents and see significant changes in the binding ability of antibodies. We have had instances where milk has prevented the binding of an antibody but where BSA gave very a good signal. Please pass on all our technical recommendations to our customers as if they do not follow our recommendations we will not offer replacements or refunds if the antibody was supposed to work as described on the datasheet. I would therefore appreciate you asking the customer to try with a different blocking agent of 5% BSA for 1hr (incubating the antibody in TBST only). If the customer still experiences problems with this modification please do not hesitate to contact us again,

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