AB257745

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

Name: Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell lysate (ab257745) | Abcam
Brand: Abcam Limited
SKU: AB257745
Price: 111000 JPY
Availability: InStock

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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.

別名を表示する

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

3 Images

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='/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='/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

false

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 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

細胞タイプ

HEK-293T

生物種

Human

組織

Kidney

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

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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製品の詳細

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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製品内容

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出荷温度及び保存条件

遺伝子名

TK1

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

出荷温度

Ambient - Can Ship with Ice

短期保存温度

-20°C

長期保存温度

-20°C

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補足情報

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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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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Abcam product promise

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