NT5E KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 121 bp deletion in exon 2.
Images
Key facts
- Cell type
- A-375
- Species or organism
- Human
- Tissue
- Skin
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 121 bp deletion in exon 2
Alternative names
5' nucleotidase (CD73), 5' nucleotidase precursor, 5' nucleotidase, ecto, 5' nucleotidase, ecto (CD73), 5'-NT, 5'-nucleotidase, 5NTD_HUMAN, CD73, CD73 antigen, E5NT, Ecto-5'-nucleotidase, NT, NT5, NT5E, Purine 5 Prime Nucleotidase, eN, eNT
What's included?
Recommended products
NT5E KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 121 bp deletion in exon 2.
Key facts
- Cell type
- A-375
- Species or organism
- Human
- Tissue
- Skin
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 121 bp deletion in exon 2
- Disease
- Melanoma
- Concentration
Properties
- Gene name
- NT5E
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Female
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
CD73 also known as ecto-5'-nucleotidase is an enzyme that functions at the cellular level. The CD73 protein hydrolyzes extracellular nucleotides into nucleosides by removing phosphate groups. It is a 70 kDa glycosylphosphatidylinositol-anchored molecule found on the surface of various cell types including endothelial cells lymphocytes and epithelial cells. Its presence is noted in a wide range of tissues across the human body highlighting its versatile role in cellular communication and metabolism.
- Biological function summary
CD73 participates in the regulation of purinergic signaling through adenosine production. It acts in cooperation with other cell surface enzymes such as CD39 forming a functional complex that handles ATP breakdown to adenosine. This enzymatic activity modulates immune responses tissue protection and inflammation control. In addition to immune-related functions CD73 supports the maintenance of vascular integrity and proper cellular adhesion which is important in different physiological and pathological contexts.
- Pathways
CD73 plays an important part in the adenosinergic pathway governing the conversion of AMP into adenosine. This activity impacts the signaling and function of the adenosine receptors on immune and non-immune cells. Another important pathway is the angiogenesis process where CD73 contributes by affecting endothelial cell migration and blood vessel development. Relationships with key proteins such as adenosine A2 receptors help facilitate these critical pathway activities.
- Associated diseases and disorders
CD73's involvement with immunosuppression and tumor progression becomes significant. Elevated levels of CD73 are frequently observed in some cancers where it aids in creating a favorable tumor microenvironment through increased adenosine production. This situation assists tumor cells in evading immune detection. Additionally CD73 is linked to autoimmune diseases like multiple sclerosis where it contributes to modulated lymphocyte activity. Connections with proteins such as TGF-beta further illustrate its role in disease-related pathways creating potential targets for therapeutic interventions through the use of anti-CD73 antibodies.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
Western blot - Human NT5E knockout A375 cell lysate (ab282943)
Lane 1: Wild-type A-375 cell lysate 20 μg
Lane 2: NT5E knockout A-375 cell lysate 20 μg
Lane 3: A431 cell lysate 20 μg
Lane 4: HepG2 cell lysate 20 μg
False colour image of Western blot: Anti-CD73 antibody [4G6E3] staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD73 antibody [4G6E3] ab202122 was shown to bind specifically to CD73. A band was observed at 65-70 kDa in wild-type A-375 cell lysates with no signal observed at this size in NT5E knockout cell line Human NT5E knockout A375 cell line ab281591 (knockout cell lysate ab282943). A cross-reactive band is observed at 65 kDa in all samples. To generate this image, wild-type and NT5E knockout A-375 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.All lanes: Western blot - Anti-CD73 antibody [4G6E3] (Anti-CD73 antibody [4G6E3] ab202122) at 1/500 dilution
Lane 1: Wild-type A-375 cell lysate at 20 µg
Lane 2: NT5E knockout A-375 cell lysate at 20 µg
Lane 2: Western blot - Human NT5E knockout A375 cell line (Human NT5E knockout A375 cell line ab281591)
Lane 3: A431 cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 63 kDa
Observed band size: 65 kDa, 65-70 kDa
Sanger Sequencing - Human NT5E knockout A375 cell lysate (ab282943)
121 bp deletion in exon 2
Western blot - Human NT5E knockout A375 cell lysate (ab282943)
All lanes: Western blot - Anti-CD73 antibody [EPR6114] (Anti-CD73 antibody [EPR6114] ab133582) at 1/1000 dilution
Lane 1: Wild-type A-375 cell lysate at 20 µg
Lane 2: NT5E knockout A-375 cell lysate at 20 µg
Lane 3: A431 cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
SecondaryLanes 1 - 4: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Lanes 1 - 4: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 70 kDa
Downloads
Product protocols
- Visit the general protocols
- Visit troubleshooting
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