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WT1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.

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Images

Next Generation Sequencing - Human WT1 knockout MCF7 cell line (AB289523), expandable thumbnail

Key facts

Cell type

MCF7

Species or organism

Human

Tissue

Breast

Form

Liquid

Knockout validation

Next Generation Sequencing

Alternative names

Recommended products

WT1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.

Key facts

Cell type

MCF7

Form

Liquid

Disease

Adenocarcinoma

Concentration
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Properties

Gene name

WT1

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Next Generation Sequencing

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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.

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

Notes

Recommended control: Human wild-type MCF7 cell line (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.

Supplementary info

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

Activity summary

The Wilms Tumor Protein commonly known as WT1 is a critical transcription factor implicated in several cellular processes. It is also identified by other names such as Wilms Tumor Suppressor and WT33. WT1 protein has a mass of approximately 52-54 kDa. It is broadly expressed in various tissues with high levels in the developing kidney gonads and certain mesothelial tissues. Its expression plays an important role in organ development and cellular differentiation.

Biological function summary

The WT1 protein regulates gene expression by binding to specific DNA sequences influencing cell growth and differentiation. WT1 acts as a part of larger protein complexes interacting with other transcription factors and co-regulators. It plays essential roles in the development of the urogenital system and in the maintenance of mesothelial cells. The WT1 protein is essential for normal kidney and gonadal development highlighting its significance in embryogenesis.

Pathways

WT1 integration involves multiple cellular signaling cascades. It is notably engaged in the Wnt and PI3K/AKT signaling pathways which are critical for cell proliferation and survival. In these pathways WT1 interaction with proteins such as β-catenin and PI3K subunits modulates cell fate decisions. This involvement in signaling networks exemplifies its importance in cellular homeostasis and response to environmental cues.

Associated diseases and disorders

WT1 mutations or dysregulation associate strongly with specific pathologies particularly Wilms tumor and acute myeloid leukemia (AML). In Wilms tumor WT1 acts as a tumor suppressor gene and its loss of function leads to tumorigenesis in the kidney. In AML aberrant WT1 expression affects normal hematopoiesis and is often linked to poor prognosis. Interaction with proteins like BSA in immunohistochemistry diagnostics highlights WT1’s significance as a biomarker for these diseases.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

1 product image

  • Next Generation Sequencing - Human WT1 knockout MCF7 cell line (ab289523), expandable thumbnail

    Next Generation Sequencing - Human WT1 knockout MCF7 cell line (ab289523)

    56 bp deletion (allele 1) and 55 bp deletion (allele 2) in exon 2, CCDS5750.1

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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