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AB247867

Anti-Nucleoside phosphorylase antibody [EPR5715] - BSA and Azide free

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Rabbit Recombinant Monoclonal Nucleoside phosphorylase antibody. Carrier free. Suitable for WB, ICC/IF and reacts with Human samples.

View Alternative Names

NP, PNP, Purine nucleoside phosphorylase, Inosine phosphorylase, Inosine-guanosine phosphorylase

1 Images
Western blot - Anti-Nucleoside phosphorylase antibody [EPR5715] - BSA and Azide free (AB247867)
  • WB

Lab

Western blot - Anti-Nucleoside phosphorylase antibody [EPR5715] - BSA and Azide free (AB247867)

This data was developed using the same antibody clone in a different buffer formulation (ab109447).

Lanes 1-3 : Merged signal (red and green). Green - ab109447 observed at 31 kDa. Red - loading control ab7291 observed at 50 kDa.

ab109447 Anti-Nucleoside phosphorylase antibody [EPR5715] was shown to specifically react with Nucleoside phosphorylase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266158 (knockout cell lysate ab257594) was used. Wild-type and Nucleoside phosphorylase knockout samples were subjected to SDS-PAGE. ab109447 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Nucleoside phosphorylase antibody [EPR5715] (<a href='/en-us/products/primary-antibodies/nucleoside-phosphorylase-antibody-epr5715-ab109447'>ab109447</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PNP knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PNP (Nucleoside phosphorylase) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-pnp-nucleoside-phosphorylase-knockout-hek-293t-cell-line-ab266158'>ab266158</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 32 kDa

Observed band size: 31 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR5715

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB, ICC/IF

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

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Product details

ab247867 is the carrier-free version of ab109447.

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Supplementary information

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

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.

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.

Product protocols

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

Target data

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).
See full target information PNP

Product promise

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For full details, please see our Terms & Conditions

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