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KLF4 KO cell line available now. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.

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Images

Sanger Sequencing - Human KLF4 knockout HeLa cell line (AB261752), expandable thumbnail
  • Sanger Sequencing - Human KLF4 knockout HeLa cell line (AB261752), expandable thumbnail

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2

Recommended products

KLF4 KO cell line available now. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.

Key facts

Cell type

HeLa

Form

Liquid

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2

Disease

Adenocarcinoma

Concentration
Loading...

Properties

Gene name

KLF4

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing

Cell culture

Biosafety level

EU: 2 US: 2

Viability

~ 80%

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

  • 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.

Culture medium

DMEM (High Glucose) + 10% FBS

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 long-term storage conditions

-196°C

Notes

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255448). 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.

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

Culture medium:  DMEM (High Glucose) + 10% FBS  

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 2x10E4 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:

• All seeding densities should be based on cell counts gained by established methods.

• A guide seeding density of 2x10E4 cells/cm2 is recommended.

• Cells should be passaged when they have achieved 80-90% confluence.

• Do not allow the cell density to exceed 7x10E4 cells/cm2.

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.

Supplementary info

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

Activity summary

KLF4 also known as Kruppel-like factor 4 acts as a transcription factor involved in the regulation of gene expression. It has a molecular weight of approximately 55-60 kDa. Researchers often find KLF4 expressed in epithelial tissues such as the skin and gastrointestinal tract. It regulates cellular processes like differentiation proliferation and apoptosis playing a role in maintaining homeostasis and cell identity.

Biological function summary

KLF4 impacts cell differentiation and proliferation through its role as a transcription factor. It functions within several cellular complexes and interacts with other transcription factors to modulate gene expression. KLF4 regulates stem cell maintenance by influencing gene networks involved in self-renewal and differentiation contributing particularly to pluripotency in embryonic stem cells.

Pathways

The transcriptional activity of KLF4 integrates into key signaling pathways like the Wnt/β-catenin and Notch pathways. Within the Wnt/β-catenin pathway KLF4 may cooperate with β-catenin and TCF/LEF transcription factors to modulate gene targets. Similarly in the Notch signaling pathway KLF4 interacts with pathway components to influence cell fate decisions. These interactions highlight KLF4's central role in orchestrating complex signaling networks.

Associated diseases and disorders

Mutations or dysregulated expression of KLF4 associates with various conditions including colorectal cancer and skin disorders like atopic dermatitis. In colorectal cancer KLF4 often interacts with other oncogenes and tumor suppressor proteins such as p53 to influence tumorigenesis. In skin disorders altered KLF4 expression can impact skin barrier function and inflammatory responses connecting it with proteins involved in immune response regulation.

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.

2 product images

  • Sanger Sequencing - Human KLF4 knockout HeLa cell line (ab261752), expandable thumbnail

    Sanger Sequencing - Human KLF4 knockout HeLa cell line (ab261752)

    Allele-2: Insertion of the selection cassette in exon 2.

  • Sanger Sequencing - Human KLF4 knockout HeLa cell line (ab261752), expandable thumbnail

    Sanger Sequencing - Human KLF4 knockout HeLa cell line (ab261752)

    Allele-1: 1 bp insertion in exon 2.

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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'.

For licensing inquiries, please contact partnerships@abcam.com