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OGA KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 1.

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Images

Western blot - Human MGEA5 (OGA) knockout HeLa cell line (AB265821), expandable thumbnail
  • Sanger Sequencing - Human MGEA5 (OGA) knockout HeLa cell line (AB265821), expandable thumbnail

Key facts

Cell type
HeLa
Species or organism
Human
Tissue
Cervix
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 1

Alternative names

Beta-N-acetylglucosaminidase, Beta-N-acetylhexosaminidase, Beta-hexosaminidase, Bifunctional protein NCOAT, HEXC, Hyaluronidase in meningioma, KIAA0679, MEA5, MGEA 5, Meningioma-expressed antigen 5, N-acetyl-beta-D-glucosaminidase, N-acetyl-beta-glucosaminidase, NCOAT, Nuclear cytoplasmic O-GlcNAcase and acetyltransferase, OGA_HUMAN, Protein O-GlcNAcase

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OGA KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 1.

Key facts

Cell type
HeLa
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 1
Antibiotic resistance
Puromycin 1µg/mL
Disease
Adenocarcinoma
Concentration
Loading...

Properties

Gene name
OGA
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous

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 HeLa cell line (ab255928). 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

MGEA5 also referred to as OGA or O-GlcNAcase serves an important mechanical role in cellular metabolism. This enzyme with a molecular mass of around 103 kDa is primarily involved in the removal of O-GlcNAc modifications from serine/threonine residues on nuclear and cytoplasmic proteins. It is expressed in various tissues but exhibits predominant activity in the brain liver and pancreas. Alternative names like "OGA stains" or "OGA b" may also be encountered in literature or research contexts.

Biological function summary

MGEA5 acts as an important modulator of protein function and regulation by cycling O-GlcNAc modifications thereby influencing processes like transcription signal transduction and apoptosis. While not part of a well-defined complex it frequently interacts with other proteins involved in glucose metabolism and the cellular stress response. Its activity works in concert with OGT (O-GlcNAc transferase) which adds O-GlcNAc maintaining a balance important for cellular function.

Pathways

MGEA5 involvement centers largely around glucose homeostasis and the insulin signaling pathway. It modulates the function of proteins like insulin receptor substrate (IRS) and other key regulators in this pathway affecting metabolic regulation and energy homeostasis. It shows a direct relationship with OGT as they reciprocate in action effects highlighting their collaborative role in cellular pathways that affect cellular signaling and survival.

Associated diseases and disorders

MGEA5 has been implicated in neurodegenerative diseases and diabetes. In Alzheimer's disease altered O-GlcNAc cycling affects tau protein homeostasis which is significant in disease pathogenesis. Likewise in diabetes MGEA5 influences insulin resistance by regulating O-GlcNAc levels on insulin signaling molecules. Dysregulation of these processes may also involve proteins such as beta-amyloid further linking MGEA5 to these pathological states.

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

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