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AB286311

Human TET2 knockout MCF7 cell line

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TET2 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.

View Alternative Names

FLJ20032, KIAA1546, MDS, Methylcytosine dioxygenase TET2, Nbla00191, Probable methylcytosine dioxygenase TET2, Protein Ayu17 449, TET2_HUMAN, Tet methylcytosine dioxygenase 2, Tet oncogene 2, Tet oncogene family member 2

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Next Generation Sequencing - Human TET2 knockout MCF7 cell line (AB286311)
  • NGS

Lab

Next Generation Sequencing - Human TET2 knockout MCF7 cell line (AB286311)

55 bp deletion in exon 1, CCDS47120.1

Key facts

Cell type

MCF7

Species or organism

Human

Tissue

Breast

Form

Liquid

form

Knockout validation

Next Generation Sequencing

Disease

Adenocarcinoma

Reactivity data

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

Product details

Recommended control: Human wild-type MCF7 cell line (ab288560). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

Properties and storage information

Gene name
TET2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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
  • Slow to trypsinise.
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 5-7x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

MEM + 10% FBS + 0.01 mg/ml bovine insulin

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.

Tet2 also known as ten-eleven translocation methylcytosine dioxygenase 2 is an enzyme involved in DNA demethylation. It has a molecular weight of approximately 224 kDa. This enzyme catalyzes the conversion of 5-methylcytosine to 5-hydroxymethylcytosine an important step in DNA demethylation processes. Tet2 expression occurs in various tissues with higher levels in hematopoietic cells. Its role in modification of DNA has made it a significant subject of study in epigenetic regulation.
Biological function summary

Tet2 participates in the regulation of gene expression through its impact on DNA methylation status. By promoting DNA demethylation Tet2 influences transcriptional networks that are vital for cell differentiation and development. The enzyme functions as part of a larger complex with cofactors including ascorbic acid which it requires for full enzymatic activity. Its actions affect the regulation of important processes such as self-renewal and differentiation in hematopoietic stem cells.

Pathways

The enzymatic activity of Tet2 plays a role in epigenetic pathways involved in hematopoiesis and immune function. In particular Tet2 interacts with pathways regulating chromatin structure and transcriptional control. It associates with proteins like TET1 and TET3 as they share roles in the regulation of 5-hydroxymethylcytosine levels. Through these mechanisms Tet2 contributes significantly to the maintenance of normal hematopoietic and immune cell function.

Alterations in Tet2 activity have been linked to hematological malignancies such as myelodysplastic syndromes and acute myeloid leukemia. Loss of function mutations in Tet2 lead to disruptions in hematopoietic differentiation contributing to disease pathology. Tet2 mutations can affect interactions with proteins such as DNMT3A another promoter of DNA methylation which further influence the disease course. Understanding Tet2's role in these disorders may offer potential therapeutic insights.

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

Product protocols

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