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AB257774

Human TYMP (Thymidine Phosphorylase) knockout HeLa cell lysate

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TYMP KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2.

View Alternative Names

Gliostatin, ECGF1, TdRPase, PD-ECGF, TP, Thymidine phosphorylase, TYMP, Platelet-derived endothelial cell growth factor

2 Images
Sanger Sequencing - Human TYMP (Thymidine Phosphorylase) knockout HeLa cell lysate (AB257774)
  • Sanger seq

Unknown

Sanger Sequencing - Human TYMP (Thymidine Phosphorylase) knockout HeLa cell lysate (AB257774)

Homozygous : 1 bp insertion in exon2

Western blot - Human TYMP (Thymidine Phosphorylase) knockout HeLa cell lysate (AB257774)
  • WB

Lab

Western blot - Human TYMP (Thymidine Phosphorylase) knockout HeLa cell lysate (AB257774)

False colour image of Western blot : Anti-TYMP antibody staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to TYMP. A band was observed at 50 kDa in wild-type HeLa cell lysates with no signal observed at this size in TYMP knockout cell line ab265138 (knockout cell lysate ab257774). To generate this image, wild-type and TYMP knockout HeLa 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-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TYMP (Thymidine Phosphorylase) knockout HeLa cell lysate (ab257774) at 20 µg

Lane 3:

THP-1 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Secondary

Lanes 1 - 4:

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 1/20000 dilution

Lanes 1 - 4:

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

false

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2.

Disease

Adenocarcinoma

Reactivity data

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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 and patent pages.

What's included?

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Properties and storage information

Gene name
TYMP
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
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.

Thymidine phosphorylase also known as platelet-derived endothelial cell growth factor (PD-ECGF) acts mechanically to catalyze the reversible phosphorolysis of thymidine into thymine and 2-deoxy-alpha-D-ribose-1-phosphate. With a mass of approximately 44 kDa this enzyme is present in various tissues including blood vessels and macrophages. Its expression tends to be higher in tumor tissues which suggests a specific role in cancer biology.
Biological function summary

The enzyme participates in pyrimidine nucleoside metabolism providing an alternate route for thymidine catabolism. It is not part of a larger complex functioning independently to promote angiogenesis. Thymidine phosphorylase enhances the degradation of extracellular thymidine which can positively impact nucleotide balance within cells aiding DNA synthesis and repair.

Pathways

Thymidine phosphorylase operates significantly within the nucleotide salvage pathways. It is essential for clarifying thymidine availability for DNA synthesis. The enzyme links with enzymes like dihydropyrimidine dehydrogenase in pyrimidine degradation playing a critical role in maintaining cellular nucleotide homeostasis.

Altered activity of thymidine phosphorylase correlates with cancer and mitochondrial neurogastrointestinal encephalomyopathy (MNGIE). In tumors increased levels of this enzyme facilitate angiogenesis contributing to tumor growth and metastasis. In MNGIE mutations in this enzyme lead to imbalances in nucleotides causing mitochondrial dysfunction. The enzyme's dysregulation connects to angiogenesis-related proteins like VEGF in cancer highlighting potential therapeutic targets.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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