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MELK KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 9 and Insertion of the selection cassette in exon 9.

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Images

Western blot - Human MELK knockout HCT116 cell line (AB266896), expandable thumbnail
  • Sanger Sequencing - Human MELK knockout HCT116 cell line (AB266896), expandable thumbnail
  • Sanger Sequencing - Human MELK knockout HCT116 cell line (AB266896), expandable thumbnail

Key facts

Cell type
HCT116
Species or organism
Human
Tissue
Colon
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 9 and Insertion of the selection cassette in exon 9

Alternative names

AI327312, HPK 38, KIAA0175, Likely ortholog of maternal embryonic leucine zipper kinase, MELK_HUMAN, MPK38, Maternal embryonic leucine zipper kinase, OTTHUMP00000021377, OTTHUMP00000046113, Protein kinase Eg3, Protein kinase PK38, RP23 382O11.1, Tyrosine protein kinase MELK, hMELK, mKIAA0175, pEg3 kinase

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MELK KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 9 and Insertion of the selection cassette in exon 9.

Key facts

Cell type
HCT116
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 9 and Insertion of the selection cassette in exon 9
Disease
Carcinoma
Concentration
Loading...

Properties

Gene name
MELK
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot

Quality control

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

Cell culture

Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male

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
McCoY5a + 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 short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type HCT116 cell line (ab255451). 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, 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

Maternal embryonic leucine zipper kinase commonly referred to as MELK is a serine/threonine protein kinase with a molecular mass of about 74 kDa. MELK is broadly expressed in both human and animal tissues with significant presence in embryonic and adult stem cells. Researchers often recognize it as MELK or MELK-0A21 in studies. The protein is characterized by its regulation of cell division and apoptosis making it an important player in cellular proliferation.

Biological function summary

MELK regulates signals that control the cell cycle apoptosis and embryonic development. MELK works as a component of complexes involved in maintaining cellular homeostasis. The protruding roles involve influencing cellular architecture and governance of stem cell maintenance. Studies in animal models show MELK involvement in maintaining pluripotency and regulation during the development phase.

Pathways

MELK participates in key processes such as the cell cycle and apoptosis pathways. Its diverse functionality connects MELK to proteins like Bcl-2-associated death promoter (BAD) and p53 which further influence main cellular events. These pathways and interactions make MELK a target of interest in the study of cellular growth and programmed cell death.

Associated diseases and disorders

Researchers link MELK to cancer and neurodegenerative diseases. The overexpression of MELK is frequently observed in various cancer types making it relevant to oncogenesis. Studies examine its relation to proteins like cyclin-dependent kinases (CDKs) and survivin in cancer progression. Additionally MELK-0A21 variants may offer insights in disorders like Glioblastoma where abnormal MELK activity aligns with disease aggression.

Product promise

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

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Product protocols

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

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