GALK1 Stable Knockout HEK293T Cell Line

T61531x106 cells / 1.0 ml


DescriptionThe Galactokinase 1 (GALK1) is major enzyme in the Leloir pathway, the primary pathway for galactose metabolism. Specifically, this enzyme modifies galactose with a phosphate group to generate galactose 1-phosphate which serves as a precursor for O-linked glycosylation. Deficiencies in GALK1 leads to Galactosemia: a congenital condition that causes galactose to accumulate in blood and tissue. GALK1 Stable Knockout cell line was generated through the knockout of the GALK1 gene in HEK293T cells using the CRISPR-Cas 9 system. Cells were transiently transfected with an expression vector containing a GFP tag and a Guide RNA sequence that targeted GALK1. Clones were initially chosen through strong GFP expression and knockout of GALK1 confirmed via sequencing and GALK1 enzymatic activity. In combination with the GALE GALK1 Stable Knockout cell line (T6151), this line may prove useful in the study of O-linked glycosylation and galactose metabolism.
SpeciesHuman (H. sapiens)
Tissue/Organ/Organ SystemKidney
Donor AgeEmbryo
Growth PropertiesAdherent
Cell MorphologyEpithelial
Seeding Density30,000 - 40,000 cells/cm2
Population Doubling Time18 - 28 hours

For Research Use Only

Unit quantity1x106 cells / 1.0 ml
Pharmaceutical TargetEnzymes
Cell TypeDrug Discovery Cell Lines
Expression Profile

Knock out of GALK1

Propagation Requirements
Use of PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.
The base medium for this cell line is Prigrow III available from abm, Cat. No. TM003. To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10% and Penicillin/Streptomycin Solution (G255).
Carbon dioxide (CO2): 5%, Temperature: 37.0

1) Confirmed GALK1 knockout through sequencing (Figure S2). Tested O-glycosylation levels through enzymatic activity on the substrate SIVmac239 gp120 via Western Blot and SDS gel analysis (Figure 4).


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2. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected].

3. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.

4. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).

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DepositorUniversity of Miami

Supporting Protocol




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