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AB265644

Human GALK1 knockout HeLa cell line

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GALK1 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 2 bp insertion in exon 1.

View Alternative Names

EC 2.7.1.6, GALK, GALK1_HUMAN, GK1, GLK1, Galactokinase, Galactokinase 1, Galactose kinase, HEL-S-19, epididymis secretory protein Li 19

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Sanger Sequencing - Human GALK1 knockout HeLa cell line (AB265644)
  • Sanger seq

Unknown

Sanger Sequencing - Human GALK1 knockout HeLa cell line (AB265644)

Allele-2 : 2 bp insertion in exon 1.

Sanger Sequencing - Human GALK1 knockout HeLa cell line (AB265644)
  • Sanger seq

Unknown

Sanger Sequencing - Human GALK1 knockout HeLa cell line (AB265644)

Allele-1 : 1 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, 1 bp deletion in exon 1 and 2 bp insertion in exon 1

Disease

Adenocarcinoma

Product details

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.

What's included?

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Properties and storage information

Gene name
GALK1
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.

GALK1 also known as galactokinase 1 is an enzyme with a molecular mass of approximately 42 kDa. It catalyzes the phosphorylation of galactose to galactose-1-phosphate an important first step in galactose metabolism. GALK1 is expressed mainly in the liver kidney and brain where it plays a critical role in processing galactose derived from the diet. Its activity ensures the proper utilization of galactose for energy production and biosynthetic processes.
Biological function summary

The phosphorylation by GALK1 initiates the Leloir pathway facilitating efficient galactose metabolism. GALK1 does not form part of a larger protein complex but acts independently to enable the conversion of galactose to intermediates useful in glycolysis and glycogenesis. By doing so it supports normal cellular functions and contributes to the maintenance of cellular energy balance.

Pathways

GALK1 functions at the beginning of the Leloir pathway critical for galactose processing. This pathway includes the conversion of galactose-1-phosphate to glucose-1-phosphate a reaction controlled by related enzymes like GALT (galactose-1-phosphate uridylyltransferase). GALK1's activity is interconnected with glucose metabolism bridging galactose and glucose metabolic pathways ensuring galactose can be converted for energy production or storage.

Mutations in GALK1 can lead to galactokinase deficiency which causes galactosemia type II. This condition results in the accumulation of galactitol leading to cataracts development in early childhood. Galactokinase deficiency can also disturb normal carbohydrate metabolism indirectly affecting other proteins such as GALT. Understanding GALK1's function and genetic variants helps in diagnosing and managing galactosemia and its related complications.

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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