Human PHKA1 knockout HEK-293T cell line
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- WB
Lab
Western blot - Human PHKA1 knockout HEK-293T cell line (AB267337)
Lanes 1 - 2:
Western blot - Anti-PHKA1 antibody [EPR12118] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/phka1-antibody-epr12118-bsa-and-azide-free-ab249913'>ab249913</a>) at 1/1000 dilution
Lanes 1 - 2:
Western blot - Anti-PHKA1 antibody [EPR12118] (<a href='/en-us/products/primary-antibodies/phka1-antibody-epr12118-ab176338'>ab176338</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
PHKA1 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human PHKA1 knockout HEK-293T cell line (ab267337)
Lane 2:
Western blot - Human PHKA1 knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-phka1-knockout-hek-293t-cell-lysate-ab258111'>ab258111</a>)
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 130 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PHKA1 knockout HEK-293T cell line (AB267337)
Homozygous : 2 bp deletion in exon14
- Cell Culture
Unknown
Cell Culture - Human PHKA1 knockout HEK-293T cell line (AB267337)
Representative images of PHKA1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHKA1 antibody [EPR12118] (AB176338)](https://content.abcam.com/products/images/phka1-antibody-epr12118-ab176338--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img54450.jpg)
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Biological function summary
Glycogen metabolism involves PHKA1 as part of the phosphorylase kinase complex. This complex exists in the muscle and liver and includes alpha beta gamma and delta subunits. PHKA1 binds together with these subunits to form a large complex helping to regulate the activity state of the enzyme. The regulation of glycogen phosphorylase is important for maintaining energy supply in cells particularly during physical activity.
Pathways
The activity of PHKA1 impacts glycogenolysis which is a significant metabolic pathway responsible for breaking down glycogen into glucose-1-phosphate. This pathway involves other proteins like glycogen phosphorylase and phosphoprotein phosphatase 1. PHKA1 has interactions within the cascade that ensures proper glycogen breakdown especially during conditions requiring increased energy output.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com