Western Blot (WB)
(Figure 1. Western blot analysis of PNP using anti-PNP antibody (MBS1753983).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: human HEK293 whole cell lysatesLane 2: human PC-3 whole cell lysatesLane 3: human U87 whole cell lysatesLane 4: human A549 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- PNP antigen affinity purified monoclonal antibody (Catalog # MBS1753983) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for PNP at approximately 32KD. The expected band size for PNP is at 32KD. )
Mouse anti-Human PNP Monoclonal Antibody | anti-PNP antibody
Anti-PNP Antibody (monoclonal, 7G5)
IHC-P: 2-5ug/ml|Human|
ICC/IF: 5ug/ml|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of PNP using anti-PNP antibody (MBS1753983).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: human HEK293 whole cell lysatesLane 2: human PC-3 whole cell lysatesLane 3: human U87 whole cell lysatesLane 4: human A549 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- PNP antigen affinity purified monoclonal antibody (Catalog # MBS1753983) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for PNP at approximately 32KD. The expected band size for PNP is at 32KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of PNP using anti-PNP antibody (MBS1753983).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: human HEK293 whole cell lysatesLane 2: human PC-3 whole cell lysatesLane 3: human U87 whole cell lysatesLane 4: human A549 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- PNP antigen affinity purified monoclonal antibody (Catalog # MBS1753983) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for PNP at approximately 32KD. The expected band size for PNP is at 32KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of PNP using anti-PNP antibody (MBS1753983).PNP was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-PNP Antibody (MBS1753983) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of PNP using anti-PNP antibody (MBS1753983).PNP was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-PNP Antibody (MBS1753983) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of PNP using anti-PNP antibody (MBS1753983).PNP was detected in paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-PNP Antibody (MBS1753983) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of PNP using anti-PNP antibody (MBS1753983).PNP was detected in paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-PNP Antibody (MBS1753983) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunofluorescence (IF)
(Figure 4. IF analysis of PNP using anti- PNP antibody (MBS1753983).PNP was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- PNP Antibody (MBS1753983) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Immunofluorescence (IF)
(Figure 4. IF analysis of PNP using anti- PNP antibody (MBS1753983).PNP was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- PNP Antibody (MBS1753983) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
2. Frecker, M., Dallaire, L., Young, S. R., Chen, G. C. C., Simpson, N. E. Confirmation of regional assignment of nucleoside phosphorylase (NP) on chromosome 14 by gene dosage studies. Hum. Genet. 45: 167-173, 1978.
3. Markert, M. L., Finkel, B. D., McLaughlin, T. M., Watson, T. J., Collard, H. R., McMahon, C. P., Andrews, L. G., Barrett, M. J., Ward, F. E. Mutations in purine nucleoside phosphorylase deficiency. Hum. Mutat. 9: 118-121, 1997.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes an enzyme which reversibly catalyzes the phosphorolysis of purine nucleosides. The enzyme is trimeric, containing three identical subunits. Mutations which result in nucleoside phosphorylase deficiency result in defective T-cell (cell-mediated) immunity but can also affect B-cell immunity and antibody responses. Neurologic disorders may also be apparent in patients with immune defects. A known polymorphism at aa position 51 that does not affect enzyme activity has been described. A pseudogene has been identified on chromosome 2. [provided by RefSeq, Jul 2008]
Uniprot Description
NP: a metabolic enzyme with purine-nucleoside phosphorylase activity. Defects are the cause of nucleoside phosphorylase deficiency (NP deficiency), with severe T-cell immunodeficiency with neurologic disorder in children.
Protein type: Nucleotide Metabolism - purine; Nucleotide Metabolism - pyrimidine; Cofactor and Vitamin Metabolism - nicotinate and nicotinamide; EC 2.4.2.1; Transferase
Chromosomal Location of Human Ortholog: 14q13.1
Cellular Component: cytoskeleton; cytoplasm; intracellular; cytosol
Molecular Function: purine binding; nucleoside binding; drug binding; phosphate binding; purine-nucleoside phosphorylase activity
Biological Process: response to drug; nicotinamide riboside catabolic process; inosine catabolic process; nucleobase, nucleoside and nucleotide metabolic process; nucleobase, nucleoside, nucleotide and nucleic acid metabolic process; purine nucleotide catabolic process; immune response; positive regulation of T cell proliferation; positive regulation of alpha-beta T cell differentiation; purine base metabolic process; purine salvage
Disease: Purine Nucleoside Phosphorylase Deficiency
Research Articles on PNP
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Product Notes
The PNP pnp (Catalog #AAA1753983) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-PNP Antibody (monoclonal, 7G5) reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's PNP can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF). WB: 0.25-0.5ug/ml|Human| IHC-P: 2-5ug/ml|Human| ICC/IF: 5ug/ml|Human|. Researchers should empirically determine the suitability of the PNP pnp for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "PNP, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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