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AB257745

Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell lysate

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TK1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.

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

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Western blot - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell lysate (AB257745)
  • WB

Lab

Western blot - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell lysate (AB257745)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : TK1 knockout HEK-293T cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab76495 observed at 25 kDa. Red - loading control ab7291 observed at 50 kDa.

ab76495 Rabbit monoclonal [EPR3193] to Thymidine Kinase 1/TK1 was shown to specifically 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 and Thymidine Kinase 1/TK1 knockout samples 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) were incubated overnight at 4°C at 1 in 1000 dilution and 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 (<a href='/en-us/products/cell-lines/human-tk1-thymidine-kinase-1-knockout-hek-293t-cell-line-ab266230'>ab266230</a>)

Predicted band size: 21 kDa,25 kDa

Observed band size: 21 kDa,25 kDa

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Sanger Sequencing - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell lysate (AB257745)
  • Sanger seq

Unknown

Sanger Sequencing - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell lysate (AB257745)

Allele-1 : Insertion of the selection cassette in exon 1

Sanger Sequencing - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell lysate (AB257745)
  • Sanger seq

Unknown

Sanger Sequencing - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell lysate (AB257745)

Allele-2 : 4 bp deletion in exon 1

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, 4 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.

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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.

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What's included?

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Properties and storage information

Gene name
TK1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Thymidine Kinase 1 (TK1) also known as Thymidine Kinase isoform A is an enzyme involved in the phosphorylation of thymidine into thymidine monophosphate an important component in DNA synthesis. TK1 has a molecular weight of approximately 25 kDa. It is predominantly expressed in proliferating cells with expression levels peaking during the S phase of the cell cycle. You often find TK1 in tissues with high cell division rates like bone marrow and lymphatic tissues.
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.

TK1 levels often increase in cancerous tissues due to their elevated cell proliferation rates. Serum TK1 is a biomarker for various cancers like leukemia and breast cancer. Studies show a correlation between high TK1 activity and tumor aggressiveness and progression. Additionally interactions of TK1 with the protein p53 a tumor suppressor show that defects in TK1 regulation can contribute to genomic instability highlighting its role in oncogenesis.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB76495

Anti-Thymidine Kinase 1/TK1 antibody [EPR3193]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (AB76495)

  • 4
  • 2
Proteins & Peptides

AB128431

Recombinant Human Thymidine Kinase 1/TK1 protein

SDS-PAGE - Recombinant Human Thymidine Kinase 1/TK1 protein (AB128431)

  • 0
  • 0
Biochemicals

AB142567

BrdU (5-bromo-2'-deoxyuridine), Thymidine analog

Chemical Structure - BrdU (5-bromo-2'-deoxyuridine), Thymidine analog (AB142567)

  • 5
  • 1
Primary Antibodies

AB185228

Anti-TREX1 antibody [EPR14985]

RabMAb|Recombinant|KO Validated

Western blot - Anti-TREX1 antibody [EPR14985] (AB185228)

  • 5
  • 1

Product promise

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