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DEGS1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 1 and 1 bp insertion in exon 1.

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Images

Western blot - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481), expandable thumbnail
  • Western blot - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481), expandable thumbnail
  • Cell Culture - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481), expandable thumbnail
  • Sanger Sequencing - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481), expandable thumbnail
  • Sanger Sequencing - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481), expandable thumbnail

Publications

Key facts

Cell type
HEK-293T
Species or organism
Human
Tissue
Kidney
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 1 and 1 bp insertion in exon 1

Alternative names

DEGS, Degenerative spermatocyte homolog 1, Degenerative spermatocyte homolog 1 lipid desaturase, Degenerative spermatocyte homolog lipid desaturase, Delta 4 desaturase sphingolipid 1, Des 1, Dihydroceramide desaturase, FADS7, MGC5079, MIG15, Membrane fatty acid (lipid) desaturase, Membrane lipid desaturase, Migration inducing gene 15 protein, Sphingolipid delta 4 desaturase, Sphingolipid delta(4) desaturase DES1

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DEGS1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 1 and 1 bp insertion in exon 1.

Key facts

Cell type
HEK-293T
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 1 and 1 bp insertion in exon 1
Concentration
Loading...

Properties

Gene name
DEGS1
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: 2 US: 2
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 short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type HEK293T cell line (ab255449). 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

MLD also known as Metachromatic Leukodystrophy protein is an enzyme of significant importance in cellular function. This protein has a molecular mass of around 50 kDa. It is primarily expressed in the brain but is also found in peripheral tissues. MLD breaks down sulfatides which are important components of myelin sheaths. The myelin sheaths encapsulate nerve cells and are essential for efficient nerve signal transmission.

Biological function summary

Metachromatic Leukodystrophy protein plays an important role in the maintenance of the nervous system structure and function. It is involved in the degradation of certain lipid compounds to prevent toxic accumulation. When the function of the MLD protein is impaired sulfatides accumulate leading to cellular damage and functional deficits. MLD does not generally function as part of a larger complex; instead it acts independently within its cellular environment.

Pathways

Metachromatic Leukodystrophy protein is integral to the sphingolipid degradation pathway. This pathway includes the catabolism of complex lipid molecules preventing harmful buildup within cells. Another enzyme arylsulfatase A closely interacts with the MLD protein and participates in the same pathway further demonstrating its importance in managing cellular lipids and maintaining cell health.

Associated diseases and disorders

Metachromatic Leukodystrophy protein is directly related to Metachromatic Leukodystrophy a genetic disorder affecting the nervous system. This disease results from mutations in the gene that encodes for the MLD protein leading to demyelination and neurological decline. Other proteins like cerebroside sulfate become dysregulated in this disorder indicating a complex network of biomolecular interactions disrupted when MLD function is compromised.

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