Description
Key facts
Cell type:HEK-293T,
Species or organism:Human,
Tissue:Kidney,
Knockout validation:Sanger Sequencing,Western blot,
Mutation description:Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
Product details:
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation:
Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10).
This means that the protein of interest is denatured.
If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions:
Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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
patent pages
Properties and Storage Information:
Gene name-ITPA, Gene editing type-Knockout, Gene editing method-CRISPR technology, Knockout validation-Sanger Sequencing, Western blot, Zygosity-Homozygous, Shipped at conditions-Ambient - Can Ship with Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
ITPA also known as inosine triphosphate pyrophosphatase is an important enzyme responsible for catalyzing the hydrolysis of inosine triphosphate (ITP) to inosine monophosphate (IMP). This enzyme prevents the accumulation of non-canonical nucleotides in the cell. ITPA has a mass of approximately 21 kDa and is expressed ubiquitously in various tissues indicating its importance across different cell types. The enzyme is also present in HEK 293T cells as researchers often use these cells for studies due to their growth characteristics and transfection efficiency.
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.
ITPA has associations with certain pathological conditions. For example ITPA deficiency can lead to inosine triphosphate accumulation which might contribute to hemolytic anemia. Mutations in the ITPA gene can also impact drug metabolism notably in patients receiving thiopurine drugs such as those used for leukemia treatment. These mutations may lead to adverse drug reactions due to the altered nucleotide balance. The connection with these conditions emphasizes the role of ITPA in maintaining cellular homeostasis and its potential influence on pharmacogenomics.
