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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.

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Images

Functional Studies - Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric) (AB204706), expandable thumbnail
  • Functional Studies - Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric) (AB204706), expandable thumbnail
  • Functional Studies - Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric) (AB204706), expandable thumbnail

Key facts

Detection method

Fluorescent

Sample types

Purified protein, Tissue Lysate, Cell Lysate

Assay type

Enzyme activity

Reactive species

Mammals

Sensitivity

= 0.005 µU

Target data

Function

Catalyzes the phosphorolytic breakdown of the N-glycosidic bond in the beta-(deoxy)ribonucleoside molecules, with the formation of the corresponding free purine bases and pentose-1-phosphate (PubMed:9305964, PubMed:23438750). Preferentially acts on 6-oxopurine nucleosides including inosine and guanosine (PubMed:9305964).

Alternative names

What's included?

100 Test
Components
10X PNP Assay Buffer
1 x 10 mL
Hypoxanthine Standard
1 x 100 µL
Inosine Substrate
1 x 200 µL
OxiRed Probe
1 x 200 µL
PNP Enzyme Mix
1 x 1 Vial
PNP Positive Control
1 x 1 Vial

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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.

Alternative names

Key facts

Detection method

Fluorescent

Sample types

Purified protein, Tissue Lysate, Cell Lysate

Assay type

Enzyme activity

Reactive species

Mammals

Assay Platform

Microplate reader

Sensitivity

= 0.005 µU

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

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.

This product is manufactured by BioVision, an Abcam company and was previously called K767 Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric). K767-100 is the same size as the 100 test size of ab204706.

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Nucleoside phosphorylase also known as PNP or purine nucleoside phosphorylase is an enzyme with a vital mechanical role in nucleoside metabolism. The enzyme facilitates the phosphorolytic cleavage of the glycosidic bond in nucleosides producing ribose 1-phosphate and free bases such as guanine and hypoxanthine. This catalytic function plays an important part in nucleotide salvage pathways. PNP has a molecular mass of approximately 32000 Daltons and shows expression predominantly in the lymphoid tissues including the thymus and spleen.

Biological function summary

Nucleoside phosphorylase is important in maintaining nucleotide homeostasis. It operates as a monomer or part of a homotrimeric complex which allows it to efficiently catalyze its reactions in purine metabolism. The absence or dysfunction of PNP results in the accumulation of nucleosides and diminished levels of nucleotide pools which can severely hamper DNA replication and repair.

Pathways

Nucleoside phosphorylase plays an integral role in the purine salvage pathway which is vital for recycling purines to form new nucleotides. It works alongside other enzymes such as adenine phosphoribosyltransferase to conserve energy by recycling purines. This pathway connects closely with the hypoxanthine-guanine phosphoribosyltransferase (HGPRT) pathway showing how PNP is intertwined in broader nucleotide biosynthesis and degradation processes.

Associated diseases and disorders

Deficiencies in nucleoside phosphorylase correlate strongly with immunodeficiencies notably purine nucleoside phosphorylase deficiency (PNP deficiency) which leads to compromised T-cell immunity. This condition can result in recurrent infections and developmental delay. Additionally the enzyme's dysregulation associates with certain leukemias where altered nucleotide pools contribute to the proliferation of malignant cells. In these contexts PNP interacts with proteins central to these disorders including those involved in purine metabolism placing it at a critical junction for potential therapeutic intervention.

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3 product images

  • Functional Studies - Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric) (ab204706), expandable thumbnail

    Functional Studies - Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric) (ab204706)

    Typical Hypoxanthine Standard calibration curve.

  • Functional Studies - Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric) (ab204706), expandable thumbnail

    Functional Studies - Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric) (ab204706)

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

  • Functional Studies - Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric) (ab204706), expandable thumbnail

    Functional Studies - Purine Nucleoside Phosphorylase Activity Assay Kit (Fluorometric) (ab204706)

    PNP specific activity in Jurkat Cell lysate.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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