Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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TK1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 1 and Insertion of the selection cassette in exon 1. 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
KITH_HUMAN, TK 1, TK 2, Thymidine kinase, Thymidine kinase 1, Thymidine kinase 1 soluble, Thymidine kinase 1 soluble isoform, Thymidine kinase cytosolic, Tk1a, Tk1b, cytosolic
- WB
Lab
Western blot - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell line (AB266230)
Lanes 1- 2 : Merged signal (red and green). Green - ab76495 observed at 25 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab76495 was shown to react with Thymidine Kinase 1/TK1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266230 (knockout cell lysate ab257745) was used. Wild-type HEK-293T and TK1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab76495 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4° at a 1 in 1000 dilution and a 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-Thymidine Kinase 1/TK1 antibody [EPR3193] (<a href='/en-us/products/primary-antibodies/thymidine-kinase-1-tk1-antibody-epr3193-ab76495'>ab76495</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
TK1 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell line (ab266230)
Predicted band size: 21 kDa,25 kDa
Observed band size: 21 kDa,25 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell line (AB266230)
Allele-1 : Insertion of the selection cassette in exon 1
- Sanger seq
Unknown
Sanger Sequencing - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell line (AB266230)
Allele-2 : 4 bp deletion 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
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
This enzyme is critical for DNA replication and repair by providing deoxythymidine triphosphate (dTTP) a deoxynucleoside triphosphate. TK1 operates as a monomer but can form a tetrameric complex in its active state which enhances its affinity for substrates. TK1 can process various substrates including thymidine and thymidine analogs such as 5-bromo-2'-deoxyuridine (BrdU) and EdU. These substrates are vital in molecular biology as they help in measuring DNA synthesis in cell proliferation studies.
Pathways
The function of TK1 is central to the salvage pathway of nucleotide synthesis. This pathway allows recycling of thymidine from degraded DNA which is particularly important for rapidly dividing cells that need to ensure a sufficient supply of nucleotides. TK1 acts alongside other enzymes such as thymidylate synthase and ribonucleotide reductase to maintain nucleotide balance within the cell ensuring proper DNA synthesis and cellular proliferation.
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
Primary Antibodies
AB76495
Anti-Thymidine Kinase 1/TK1 antibody [EPR3193]
BOND RX™ Validated|RabMAb|Recombinant|KO Validated
![Flow Cytometry (Intracellular) - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (AB76495)](https://content.abcam.com/products/images/thymidine-kinase-1-tk1-antibody-epr3193-ab76495--flow-cytometry-intracellular-img127164.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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