Human PNP (Nucleoside phosphorylase) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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PNP KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
FLJ94043, FLJ97288, FLJ97312, Inosine phosphorylase, Inosine-guanosine phosphorylase, MGC117396, MGC125915, MGC125916, NP, NP 1, Nucleoside phosphorylase, PNP, PNP 1, PNPH_HUMAN, PRO1837, PUNP, Purine nucleoside orthophosphate ribosyltransferase, Purine nucleoside phosphorylase, Purine nucleoside phosphorylase 5a
- WB
Lab
Western blot - Human PNP (Nucleoside phosphorylase) knockout HEK-293T cell line (AB266158)
Lanes 1-4 : Merged signal (red and green). Green - ab109559 observed at 31 kDa. Red - loading control ab7291 observed at 50 kDa.
ab109559 Anti-Nucleoside phosphorylase antibody [EPR5714] 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. ab109559 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 [EPR5714] (<a href='/en-us/products/primary-antibodies/nucleoside-phosphorylase-antibody-epr5714-ab109559'>ab109559</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 (ab266158)
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
JAR 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
- WB
Lab
Western blot - Human PNP (Nucleoside phosphorylase) knockout HEK-293T cell line (AB266158)
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 (ab266158)
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
- Sanger seq
Unknown
Sanger Sequencing - Human PNP (Nucleoside phosphorylase) knockout HEK-293T cell line (AB266158)
Homozygous : 1 bp insertion in exon 2
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
D7S820, D5S818, TH01, D16S539, TPOX, CSF1PO, D13S317
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB259606
Anti-Nucleoside phosphorylase antibody [EPR22045-274] - BSA and Azide free (Detector)
RabMAb|Recombinant
![Sandwich ELISA - Anti-Nucleoside phosphorylase antibody [EPR22045-274] - BSA and Azide free (Detector) (AB259606)](https://content.abcam.com/products/images/nucleoside-phosphorylase-antibody-epr22045-274-bsa-and-azide-free-detector-ab259606--sandwich-elisa-img52299.png)
- 0
- 0
Assay Kits
AB260069
Human PNP ELISA Kit (Purine Nucleoside Phosphorylase)
Recombinant|SimpleStep
- 0
- 0
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com