Human ITPA knockout HEK-293T cell line
- Advanced Validation
- What is this?
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ITPA 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 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
C20orf37, HLC14-06-P, ITPA_HUMAN, ITPase, Inosine triphosphatase, Inosine triphosphatase (nucleoside triphosphate pyrophosphatase), Inosine triphosphate pyrophosphatase, Inosine triphosphate pyrophosphohydrolase, My049, My049 protein, NTPase, Non canonical purine NTP pyrophosphatase, Non standard purine NTP pyrophosphatase, Nucleoside triphosphate pyrophosphatase, OK/SW-cl.9, OTTHUMP00000030094, OTTHUMP00000160459, Putative oncogene protein hlc14-06-p, dJ794I6.3, inosine triphosphatase-A, nucleoside triphosphate diphosphatase
- WB
Lab
Western blot - Human ITPA knockout HEK-293T cell line (AB266679)
Lanes 1-3 : Merged signal (red and green). Green - ab134937 observed at 21 kDa. Red - loading control ab8245 observed at 36 kDa.
ab134937 Anti-ITPA antibody [EPR8780] was shown to specifically react with ITPA in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266679 (knockout cell lysate ab258477) was used. Wild-type and ITPA knockout samples were subjected to SDS-PAGE. ab134937 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-ITPA antibody [EPR8780] (<a href='/en-us/products/primary-antibodies/itpa-antibody-epr8780-ab134937'>ab134937</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ITPA knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-itpa-knockout-hek-293t-cell-lysate-ab258477'>ab258477</a>) at 20 µg
Lane 3:
Jurkat cell lysate at 20 µg
Secondary
Lanes 1 - 3:
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
Lanes 1 - 3:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 21 kDa
Observed band size: 36 kDa,21 kDa
false
- WB
Lab
Western blot - Human ITPA knockout HEK-293T cell line (AB266679)
Lanes 1-3 : Merged signal (red and green). Green - ab150420 observed at 21 kDa. Red - loading control ab8245 observed at 36 kDa.
ab150420 Anti-ITPA antibody [EPR8779] was shown to specifically react with ITPA in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266679 (knockout cell lysate ab258477) was used. Wild-type and ITPA knockout samples were subjected to SDS-PAGE. ab150420 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-ITPA antibody [EPR8779] (<a href='/en-us/products/primary-antibodies/itpa-antibody-epr8779-ab150420'>ab150420</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ITPA knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ITPA knockout HEK-293T cell line (ab266679)
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: 21 kDa
Observed band size: 21 kDa
false
- Cell Culture
Unknown
Cell Culture - Human ITPA knockout HEK-293T cell line (AB266679)
Representative images of ITPA knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human ITPA knockout HEK-293T cell line (AB266679)
Homozygous : 1 bp insertion in exon 1
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
The enzyme plays a pivotal role in nucleotide metabolism by regulating the levels of ITP deoxyinosine triphosphate (dITP) and xanthosine triphosphate (XTP). By hydrolyzing these non-standard nucleotides ITPA maintains genomic integrity and prevents mutations during DNA replication. It is not typically considered part of a larger complex but acts independently to safeguard the nucleotide pool from potentially harmful nucleotide analogs.
Pathways
ITPA's activity is essential in the purine metabolism pathway where it helps maintain a proper balance of nucleotide synthesis and degradation. This function helps prevent the incorporation of non-standard bases into DNA and RNA which can lead to genetic instability. ITPA activity is also intertwined with the salvage pathway of purine nucleotides. This connection highlights its indirect relationship with proteins involved in nucleotide synthesis and repair such as DNA polymerases and other nucleotide-metabolizing enzymes.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ITPA antibody [EPR8780] (AB134937)](https://content.abcam.com/products/images/itpa-antibody-epr8780-ab134937--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img127928.jpg)
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