GALK2+GALE Stable Knockout HEK293T Cell Line

T61521x106 cells / 1.0 ml


DescriptionThe UDP-galactose-4 epimerase (encoded by GALE) and Galactokinase 2 (GALK2) are both major enzymes in the Leloir pathway, the primary pathway for galactose metabolism. UDP-galactose-4 epimerase modifies galactose and acetyl galactose, generating precursors for O-linked glycolsylation. Deficiencies in these genes can lead to Galactosemia: a condition that is caused by the accumulation of galactose in the blood and tissue of individuals that cannot properly metabolize the galactose in their diets. As this cell line is derived from a human source, it may be useful in the study of glycosylation and how it is affected during instances of disease.
This cell line was generated by knocking out GALK2 in the GALE Stable Knockout HEK293T cell line (T6150) 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 GALK2. Clones were initially chosen through strong GFP expression and knockout of GALK2 confirmed via sequencing and GALK2 enzymatic activity.
SpeciesHuman (H. sapiens)
Tissue/Organ/Organ SystemKidney
Donor AgeEmbryo
Growth PropertiesAdherent
Cell MorphologyEpithelial
Seeding Density20,000 - 40,000 cells/cm2
Population Doubling Time22 - 32 hours

For Research Use Only

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

Knock out of GALE and GALK2 expression

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 GALK 2 knockout through Western Blot analysis (Figure 5A) and sequencing (Figure S3). Tested O-glycosylation levels through enzymatic activity on the substrate SIVmac239 gp120 via Western Blot and SDS gel analysis (Figure 5B).


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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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