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Human PNP ELISA Kit (Purine Nucleoside Phosphorylase) is a single-wash 90-min Simplestep used to quantify Human PNP (Purine Nucleoside Phosphorylase) with a sensitivity of 40 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

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Images

Sandwich ELISA - Human PNP ELISA Kit (Purine Nucleoside Phosphorylase) (AB260069), expandable thumbnail
  • Sandwich ELISA - Human PNP ELISA Kit (Purine Nucleoside Phosphorylase) (AB260069), expandable thumbnail

Key facts

Detection method
Colorimetric
Sample types
Heparin Plasma, Citrate plasma, Serum, EDTA Plasma
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
78.13 - 5000 pg/mL
Assay time
1h 30m
Sensitivity
= 40 pg/mL

Reactivity data

Application
sELISA
Reactivity
Reacts
Dilution info
-
Notes

-

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:23438750, PubMed:9305964). Preferentially acts on 6-oxopurine nucleosides including inosine and guanosine (PubMed:9305964).

Alternative names

What's included?

1 x 96 Tests
Components
10X Human PNP (Purine Nucleoside Phosphorylase) Capture Antibody
1 x 600 µL
10X Human PNP (Purine Nucleoside Phosphorylase) Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
Antibody Diluent 5BR
1 x 6 mL
Human PNP (Purine Nucleoside Phosphorylase) Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NS (ab193972)
1 x 50 mL
SimpleStep Pre-Coated 96-Well Microplate (ab206978)
1 x 1 Unit
Stop Solution
1 x 12 mL
TMB Development Solution
1 x 12 mL

Recommended products

Human PNP ELISA Kit (Purine Nucleoside Phosphorylase) is a single-wash 90-min Simplestep used to quantify Human PNP (Purine Nucleoside Phosphorylase) with a sensitivity of 40 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

Key facts

Detection method
Colorimetric
Sample types
Heparin Plasma, Citrate plasma, Serum, EDTA Plasma
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
78.13 - 5000 pg/mL
Assay time
1h 30m
Assay Platform
Pre-coated microplate (12 x 8 well strips)
Sensitivity
= 40 pg/mL

Precision

Intra assay

Sample
Serum
n
8
mean
-
SD
-
C.V.
8.3

Inter assay

Sample
Serum
n
3
mean
-
SD
-
C.V.
4.1

Recovery

Sample specific recovery

Sample type
Serum
Average %
= 96
Range
85 - 108 %
Sample type
EDTA Plasma
Average %
= 96
Range
85 - 110 %
Sample type
Heparin Plasma
Average %
= 90
Range
82 - 104 %
Sample type
Citrate plasma
Average %
= 103
Range
90 - 114 %

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Notes

Human PNP ELISA Kit (Purine Nucleoside Phosphorylase) (ab260069) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of PNP (Purine Nucleoside Phosphorylase) protein in cit plasma, edta plasma, hep plasma, and serum. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human PNP (Purine Nucleoside Phosphorylase) with 40 pg/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

Purine Nucleoside Phosphorylase (PNP) is a catabolic enzyme which breaks down purine residues by releasing the nucleobase and phosphorylating the ribose residue. It is also used to produce nucleotide monobases as a salvage pathway. PNP plays an important role in the maintenance of T-cells. This kit targets the full-length protein.

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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2 product images

  • Sandwich ELISA - Human PNP ELISA Kit (Purine Nucleoside Phosphorylase) (ab260069), expandable thumbnail

    Sandwich ELISA - Human PNP ELISA Kit (Purine Nucleoside Phosphorylase) (ab260069)

    Example of human PNP standard curve in Sample Diluent NS.

    The PNP standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • Sandwich ELISA - Human PNP ELISA Kit (Purine Nucleoside Phosphorylase) (ab260069), expandable thumbnail

    Sandwich ELISA - Human PNP ELISA Kit (Purine Nucleoside Phosphorylase) (ab260069)

    Interpolated concentrations of native PNP in human serum and plasma samples.

    The concentrations of PNP were measured in duplicate, interpolated from the PNP standard curves, and corrected for sample dilution. Undiluted samples are as follows: serum 100%, plasma (citrate) 100%, plasma (heparin) 100% and plasma (edta) 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PNP concentration was determined to be 1435.64 pg/mL in serum, 4419.64 pg/mL in plasma (citrate), 2483.57 pg/mL in plasma (heparin), and 1750.71 pg/mL in plasma (edta).

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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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