AB266230

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

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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 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

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 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='/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 Sequencing - Human TK1 (Thymidine Kinase 1) knockout HEK-293T cell line (AB266230)

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.

Key facts

細胞タイプ

HEK-293T

生物種

Human

組織

Kidney

製品の状態

Liquid

form

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

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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製品の詳細

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.

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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

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

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取り扱い方法

初回取り扱いガイドライン

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.

継代培養ガイドライン

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.

培養培地

DMEM (High Glucose) + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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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 line