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AB265001

Human NLK knockout HeLa cell line

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NLK KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 14 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'.
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Sanger Sequencing - Human NLK knockout HeLa cell line (AB265001)
  • Sanger seq

Unknown

Sanger Sequencing - Human NLK knockout HeLa cell line (AB265001)

Allele-2 : Insertion of the selection cassette in exon 1.

Sanger Sequencing - Human NLK knockout HeLa cell line (AB265001)
  • Sanger seq

Unknown

Sanger Sequencing - Human NLK knockout HeLa cell line (AB265001)

Allele-1 : 14 bp deletion in exon 1.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 1 and Insertion of the selection cassette in exon 1

Disease

Adenocarcinoma

Product details

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.

What's included?

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Properties and storage information

Gene name
NLK
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger 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
  • 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.

Supplementary information

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

NLK (Nemo-Like Kinase) functions as a serine/threonine protein kinase involved in various signaling pathways. It alternatively is known as nemo-like protein or nlk. The molecular mass of NLK is approximately 54 kDa. Expression of NLK occurs in various tissues but it is highly observed in the brain and testis. This wide expression suggests that NLK may have diverse functional roles.
Biological function summary

NLK participates in regulating cellular processes such as cell cycle apoptosis and transcription. It is not part of a larger complex but works within signaling pathways to influence these processes. NLK modulates the activity of several transcription factors particularly in the Wnt/beta-catenin signaling pathway where it negatively regulates the signaling to control gene expression relevant to cell proliferation and differentiation.

Pathways

NLK is an important player in the Wnt/beta-catenin and MAPK pathways. In the Wnt/beta-catenin pathway it associates with proteins such as TCF/LEF and beta-catenin acting to modulate transcription by phosphorylating these molecules. The MAPK pathway involves NLK in response to stress signals coordinating with proteins like TAK1 to mediate cellular responses. These pathways illustrate NLK's role in transmitting signals that govern cell fate and stress responses.

NLK has links to cancer and neurodegenerative diseases. Alterations in NLK expression or function can contribute to the development and progression of cancers by affecting the regulation of cell proliferation and apoptosis. Specifically dysregulation in the Wnt/beta-catenin pathway in which NLK acts relates to colorectal cancers. In the context of neurodegenerative diseases NLK has connections with the protein TDP-43 which is implicated in diseases such as amyotrophic lateral sclerosis highlighting its role in neuronal health and potential links to neuronal degradation.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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