Western Blot (WB)
(Figure 1. Western blot analysis of PNP using anti-PNP antibody (MBS178628).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 50ug of sample under reducing conditions.Lane 1: rat thymus tissue lysates,Lane 2: rat ovary tissue lysates,Lane 3: mouse liver tissue lysates,Lane 4: human placneta tissue 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 rabbit anti-PNP antigen affinity purified polyclonal antibody at 0.5ug/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-rabbit 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 with Tanon 5200 system. A specific band was detected for PNP at approximately 38KD. The expected band size for PNP is at 38KD. )
Rabbit PNP Polyclonal Antibody | anti-PNP antibody
Anti-PNP Antibody
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Immunohistochemistry(IHC) Paraffin: 0.5-1ug/ml
By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections.
Other applications have not been tested.
Western Blot (WB)
(Figure 1. Western blot analysis of PNP using anti-PNP antibody (MBS178628).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 50ug of sample under reducing conditions.Lane 1: rat thymus tissue lysates,Lane 2: rat ovary tissue lysates,Lane 3: mouse liver tissue lysates,Lane 4: human placneta tissue 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 rabbit anti-PNP antigen affinity purified polyclonal antibody at 0.5ug/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-rabbit 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 with Tanon 5200 system. A specific band was detected for PNP at approximately 38KD. The expected band size for PNP is at 38KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of PNP using anti-PNP antibody (MBS178628).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 50ug of sample under reducing conditions.Lane 1: rat thymus tissue lysates,Lane 2: rat ovary tissue lysates,Lane 3: mouse liver tissue lysates,Lane 4: human placneta tissue 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 rabbit anti-PNP antigen affinity purified polyclonal antibody at 0.5ug/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-rabbit 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 with Tanon 5200 system. A specific band was detected for PNP at approximately 38KD. The expected band size for PNP is at 38KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of PNP using anti-PNP antibody (MBS178628).PNP was detected in paraffin-embedded section of mouse kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) overnight at 4 degree C. Biotinylated goat anti-rabbit 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of PNP using anti-PNP antibody (MBS178628).PNP was detected in paraffin-embedded section of mouse kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) overnight at 4 degree C. Biotinylated goat anti-rabbit 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of PNP using anti-PNP antibody (MBS178628).PNP was detected in paraffin-embedded section of rat kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) overnight at 4 degree C. Biotinylated goat anti-rabbit 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of PNP using anti-PNP antibody (MBS178628).PNP was detected in paraffin-embedded section of rat kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) overnight at 4 degree C. Biotinylated goat anti-rabbit 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of PNP using anti-PNP antibody (MBS178628).PNP was detected in paraffin-embedded section of human liver cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) overnight at 4 degree C. Biotinylated goat anti-rabbit 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of PNP using anti-PNP antibody (MBS178628).PNP was detected in paraffin-embedded section of human liver cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) overnight at 4 degree C. Biotinylated goat anti-rabbit 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of PNP using anti-PNP antibody (MBS178628).PNP was detected in paraffin-embedded section of human renal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) overnight at 4 degree C. Biotinylated goat anti-rabbit 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of PNP using anti-PNP antibody (MBS178628).PNP was detected in paraffin-embedded section of human renal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) overnight at 4 degree C. Biotinylated goat anti-rabbit 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) with DAB as the chromogen. )
Flow Cytometry (FC/FACS)
(Figure 6. Flow Cytometry analysis of U937 cells using anti-PNP antibody (MBS178628).Overlay histogram showing U937 cells stained with MBS178628 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PNP Antibody (MBS178628,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight ®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 6. Flow Cytometry analysis of U937 cells using anti-PNP antibody (MBS178628).Overlay histogram showing U937 cells stained with MBS178628 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PNP Antibody (MBS178628,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight ®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 7. Flow Cytometry analysis of U251 cells using anti-PNP antibody (MBS178628).Overlay histogram showing U251 cells stained with MBS178628 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PNP Antibody (MBS178628,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight ®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 7. Flow Cytometry analysis of U251 cells using anti-PNP antibody (MBS178628).Overlay histogram showing U251 cells stained with MBS178628 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PNP Antibody (MBS178628,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight ®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Background: The PNP gene encodes purine nucleoside phosphorylase, an enzyme that catalyzes the reversible phosphorolysis of the purine nucleosides and deoxynucleosides inosine, guanosine, deoxyinosine, and deoxyguanosine. It is presented results from gene dosage studies consistent with assignment of the PNP locus to band 14q13. PNP is expressed in most tissues, with markedly greater expression in lymphoid tissues. Genetic deficiencies of PNP result in severely compromised T-lymphocyte function and neurologic dysfunction.
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: Cofactor and Vitamin Metabolism - nicotinate and nicotinamide; EC 2.4.2.1; Nucleotide Metabolism - purine; Nucleotide Metabolism - pyrimidine; Transferase
Chromosomal Location of Human Ortholog: 14q11.2
Cellular Component: cytoplasm; cytosol; intracellular
Molecular Function: drug binding; nucleoside binding; phosphate binding; purine binding; purine-nucleoside phosphorylase activity
Biological Process: immune response; inosine catabolic process; nicotinamide riboside catabolic process; nucleobase, nucleoside, nucleotide and nucleic acid metabolic process; positive regulation of alpha-beta T cell differentiation; positive regulation of T cell proliferation; purine nucleotide catabolic process; purine salvage; response to drug
Disease: Purine Nucleoside Phosphorylase Deficiency
Research Articles on PNP
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Product Notes
The PNP pnp (Catalog #AAA178628) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-PNP Antibody reacts with Human, Mouse, Rat 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 (IHC) Paraffin. Western Blot: 0.1-0.5ug/ml Immunohistochemistry(IHC) Paraffin: 0.5-1ug/ml. 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, Polyclonal 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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