Adenosylhomocysteinase (AHCY) Activity Assay Kit (Fluorometric) (ab197002)

Overview

  • Product name

    Adenosylhomocysteinase (AHCY) Activity Assay Kit (Fluorometric)
    See all Adenosylhomocysteinase kits
  • Detection method

    Fluorescent
  • Sample type

    Cell Lysate, Purified protein, Tissue Lysate
  • Assay type

    Enzyme activity
  • Sensitivity

    > 1 µU
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Adenosylhomocysteinase Activity Assay Kit (ab197002) can kinetically measure AHCY activity by detecting adenosine generation resulting from the hydrolysis of SAH. Adenosine is detected via a multi-step reaction, resulting in the generation of an intermediate that reacts with the OxiRed Probe. The fluorescent product is measured at Ex/Em = 535/587 nm. Limit of quantification (L.O.Q) is 1 µU recombinant Human AHCY.

  • Notes

    Adenosylhomocysteinase (AHCY) (EC 3.3.1.1) or S-adenosylhomocysteine hydrolase (SAHH) is an enzyme that catalyzes the reversible hydrolysis of S-Adenosyl Homocysteine (SAH) to adenosine and homocysteine. AHCY regulates the intracellular SAH concentration which in turn regulates S-adenosyl methionine (SAM)-dependent methyltransferases. Down-regulation of AHCY has been associated with certain forms of cancer and Huntington’s disease, while in Wilson’s disease; the enzyme is inhibited by the accumulated copper. Mutations in the AHCY gene cause SAHH deficiency disease.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    Adenosine Standard (10 mM) 1 x 100µl
    AHCY Assay Buffer 1 x 25ml
    AHCY Enzyme (Positive Control) 1 x 10µl
    AHCY Enzyme Mix I 1 vial
    AHCY Enzyme Mix II 1 vial
    Homogenization Buffer 1 x 60ml
    OxiRed Probe in DMSO Red 1 x 200µl
    SAH Substrate 1 x 100µl
  • Function

    Adenosylhomocysteine is a competitive inhibitor of S-adenosyl-L-methionine-dependent methyl transferase reactions; therefore adenosylhomocysteinase may play a key role in the control of methylations via regulation of the intracellular concentration of adenosylhomocysteine.
  • Pathway

    Amino-acid biosynthesis; L-homocysteine biosynthesis; L-homocysteine from S-adenosyl-L-homocysteine: step 1/1.
  • Involvement in disease

    Hypermethioninemia with S-adenosylhomocysteine hydrolase deficiency
  • Sequence similarities

    Belongs to the adenosylhomocysteinase family.
  • Cellular localization

    Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Alternative names

    • Adenosylhomocysteinase
    • AdoHcyase
    • ahcY
    • S-adenosyl-L-homocysteine hydrolase
    • SAHH_HUMAN
    see all

Images

  • Typical Adenosine Standard Curve obtained following assay protocol.

  • AHCY activity in rat kidney lysate (24 µg), HeLa cell lysate (7.6 µg) and Positive Control; S: Sample, B: Background.

  • AHCY specific activity in HeLa cell lysate (4 µg) and rat kidney lysate (28 µg). 

Protocols

References

This product has been referenced in:

  • Fu Y  et al. LncRNA H19 interacts with S-adenosylhomocysteine hydrolase to regulate LINE-1 Methylation in human lung-derived cells exposed to Benzo[a]pyrene. Chemosphere 207:84-90 (2018). Human . Read more (PubMed: 29772428) »
See 1 Publication for this product

Customer reviews and Q&As

AHCY activity in murine pancreatic tissue

Good Excellent 5/5 (Ease of Use)
Abreviews
Samples: murine pancreatic tissue (about 2*1*1 mm).
Snap freeze, thaw on ice and prepare the sample according to the manufacturer´s protocol in 300 ul lysis buffer, shredder at 20000 rpm, 6 sec. For the AHCY assay 5 ul of samples were used.
RFUs were normalized to the whole protein content.

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Submitted Jul 07 2017

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