Overview

  • Product name

    Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric)
    See all Nucleoside phosphorylase kits
  • Detection method

    Fluorescent
  • Sample type

    Cell Lysate, Purified protein, Tissue Lysate
  • Assay type

    Enzyme activity
  • Sensitivity

    0.005 µU
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric) (ab204706) is an assay where hypoxanthine formed from the breakdown of inosine is detected via a multi-step reaction, resulting in the generation of an intermediate that reacts with the PNP Probe. The fluorescent product is measured at Ex/Em = 535/587 nm. Limit of quantification is 0.005 µU recombinant Purine Nucleoside Phosphorylase.

  • Notes

    Purine Nucleoside Phosphorylase (PNP, E.C. 2.4.2.1.) is an enzyme involved in purine metabolism and it catalyzes the cleavage of the glycosidic bond of ribo- or deoxyribonucleosides, in the presence of inorganic phosphate as a second substrate, to generate the purine base and ribose-1-phosphate or deoxyribose-1-phosphate. It is one of the enzymes of the nucleotide salvage pathways that allows the cell to produce nucleotide monophosphates when the de novo synthesis pathway has been interrupted or is non-existent (as is the case in the brain). PNP is a cytosolic enzyme. PNP deficiency disease leads to toxic buildup of deoxyguanosine in T-cells leading to T-lymphocytopenia with no apparent effects on B-lymphocyte function. Inhibition of PNP is an important target for chemotherapeutic applications and treatment of T- cell mediated autoimmune diseases. PNP deficiency is also associated with neurological problems.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    Enzyme Mix 1 vial
    Hypoxanthine Standard (10 mM) 1 x 100µl
    Inosine Substrate 1 x 200µl
    PNP Assay Buffer (10x) 1 x 10ml
    PNP Positive Control 1 vial
    PNP Probe (in dry DMSO) 1 x 200µl
  • Research areas

  • Involvement in disease

    Defects in PNP are the cause of purine nucleoside phosphorylase deficiency (PNP deficiency) [MIM:613179]. It leads to a severe T-cell immunodeficiency with neurologic disorder in children.
  • Sequence similarities

    Belongs to the PNP/MTAP phosphorylase family.
  • Cellular localization

    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Alternative names

    • FLJ94043
    • FLJ97288
    • FLJ97312
    • Inosine phosphorylase
    • Inosine-guanosine phosphorylase
    • MGC117396
    • MGC125915
    • MGC125916
    • NP
    • Np1
    • Nucleoside phosphorylase
    • PNP
    • Pnp1
    • PNPH_HUMAN
    • PRO1837
    • PUNP
    • Purine nucleoside orthophosphate ribosyltransferase
    • Purine nucleoside phosphorylase
    • Purine nucleoside phosphorylase 5a
    see all

Images

  • Typical Hypoxanthine Standard calibration curve.

  • Purine Nucleoside Phosphorylase Activity in Jurkat Cell (T-lymphocyte) lysate (315 ng) and Positive Control; BG: Background.

  • PNP specific activity in Jurkat Cell lysate.

Protocols

References

ab204706 has not yet been referenced specifically in any publications.

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