GALK2+GALE Stable Knockout HEK293T Cell Line
Cat. No.
T6152
Unit
1x10⁶ cells / 1 ml
Price
Inquiry
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Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing institution.
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Specifications
| Description |
The 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. |
| Cat. No. | T6152 |
| Name | GALK2+GALE Stable Knockout HEK293T Cell Line |
| Unit | 1x10⁶ cells / 1 ml |
| Price | Inquiry |
| Category | CRISPR Stable Knockout Cell Line |
| Cell Type | CRISPR Knockout Cell Line |
| Organism | Human (H. sapiens) |
| Tissue | Kidney |
| Donor History | Embryo |
| Cell Morphology | Epithelial |
| Growth Properties | Adherent, epithelial |
| Seeding Density (cells/cm2) | 20,000 - 40,000 |
| Population Doubling Time | 24 - 34 |
| Expression Profile |
Knock out of GALE and GALK2 expression |
| Propagation Method |
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
|
| Growth Conditions | Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow III (TM003) + 10% FBS + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂ |
| Subculture Protocol |
Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent. 1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions. |
| QC |
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). |
| Application | Research Use Only. |
| Storage Condition | Vapor phase of liquid nitrogen, or below -130°C. |
| BioSafety | II |
| Disclaimer |
1. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each cell line. If you have any questions regarding this, please contact us at licensing@abmgood.com. 2. All test parameters provided in the CoA are conducted using abm's standardized culture system and 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. 3. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of order placement. 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. 4. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic or any other non-RUO application(s). 5. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature are provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 6. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period". |
| Depositor | University of Miami |
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