Human TYMP (Thymidine Phosphorylase) knockout HeLa cell line
- Advanced Validation
- What is this?
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TYMP KO cell line available to order. KO validated by. 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
ECGF, ECGF 1, Endothelial cell growth factor 1, Endothelial cell growth factor 1 platelet derived, Endothelial cell growth factor, platelet-derived, Gliostatin, MEDPS1, MNGIE, MTDPS1, PD-ECGF, PDEGF, Platelet derived endothelial growth factor, Platelet-derived endothelial cell growth factor, TP, TYPH_HUMAN, TdRPase, Thymidine phosphorylase, Tymp, hPD ECGF
- WB
Lab
Western blot - Human TYMP (Thymidine Phosphorylase) knockout HeLa cell line (AB265138)
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 (<a href='/en-us/products/cell-lysates/human-tymp-thymidine-phosphorylase-knockout-hela-cell-lysate-ab257774'>ab257774</a>) 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
- Sanger seq
Unknown
Sanger Sequencing - Human TYMP (Thymidine Phosphorylase) knockout HeLa cell line (AB265138)
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB284861
Anti-Thymidine Phosphorylase antibody [EPR25629-129]
BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|Trial Size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thymidine Phosphorylase antibody [EPR25629-129] (AB284861)](https://content.abcam.com/products/images/thymidine-phosphorylase-antibody-epr25629-129-ab284861--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img176700.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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