PCK1 KO cell line available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout.
Images
Key facts
- Cell type
A549
- Species or organism
Human
- Tissue
Lung
- Form
Liquid
- Knockout validation
Next Generation Sequencing, Western blot
Knockout.
Alternative names
GTP, PCK1, PCKGC_HUMAN, PEP carboxykinase, PEPCK-C, PEPCK1, Phosphoenolpyruvate carboxykinase, Phosphoenolpyruvate carboxykinase 1, Phosphoenolpyruvate carboxykinase 1 (soluble), Phosphoenolpyruvate carboxykinase, cytosolic, Phosphoenolpyruvate carboxykinase, cytosolic [GTP], Phosphoenolpyruvate carboxylase, Phosphopyruvate carboxylase, cytosolic [GTP]
Recommended products
PCK1 KO cell line available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout.
Alternative names
GTP, PCK1, PCKGC_HUMAN, PEP carboxykinase, PEPCK-C, PEPCK1, Phosphoenolpyruvate carboxykinase, Phosphoenolpyruvate carboxykinase 1, Phosphoenolpyruvate carboxykinase 1 (soluble), Phosphoenolpyruvate carboxykinase, cytosolic, Phosphoenolpyruvate carboxykinase, cytosolic [GTP], Phosphoenolpyruvate carboxylase, Phosphopyruvate carboxylase, cytosolic [GTP]
Key facts
- Cell type
A549
- Species or organism
Human
- Tissue
Lung
- Form
Liquid
- Knockout validation
Next Generation Sequencing, Western blot
- Mutation description
Knockout.
- Disease
Carcinoma
- Concentration
Properties
- Gene name
PCK1
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Next Generation Sequencing, Western blot
Cell culture
- Biosafety level
EU: 1 US: 1
- Adherent/suspension
Adherent
- Gender
Male
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
- Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
Dry Ice
- Appropriate long-term storage conditions
-196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type A549 cell line (Human wild-type A549 cell line ab288558). 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 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.
Supplementary info
- Activity summary
The protein PCK1 also known as phosphoenolpyruvate carboxykinase PEPC or PEPCK plays an important role in gluconeogenesis by converting oxaloacetate to phosphoenolpyruvate (PEP). It has a molecular mass of around 69 kDa. PCK1 is primarily expressed in the liver kidney and adipose tissues. Through its enzymatic activity PCK1 helps to regulate blood glucose levels allowing for the production of glucose from non-carbohydrate sources.
- Biological function summary
PCK1 is essential for maintaining glucose homeostasis in the body. Although not part of a larger protein complex its function directly impacts the tricarboxylic acid (TCA) cycle by providing substrates for glucose synthesis. By orchestrating the conversion of oxaloacetate to PEP PCK1 contributes significantly to reducing dependency on dietary carbohydrates particularly during fasting or intensive physical activity.
- Pathways
PCK1 integrates critical metabolic processes. It mainly influences the gluconeogenesis and glycolysis pathways. In the gluconeogenesis pathway PCK1 coordinates with enzymes such as glucose-6-phosphatase to facilitate the generation of glucose from lactate and amino acids. Additionally its role in glycolysis intersects with enzymes like pyruvate kinase managing energy production and consumption.
- Associated diseases and disorders
PCK1 links to conditions such as diabetes mellitus and metabolic syndrome. Altered expression or function of PCK1 can lead to disturbances in glucose metabolism. In diabetes dysregulation of PCK1 expression can contribute to hyperglycemia. Additionally PCK1 and its interaction with insulin signaling pathways is critical as insulin resistance is an important feature of metabolic syndrome. Understanding these connections offers potential avenues for therapeutic intervention in metabolic diseases.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
2 product images
Western blot - Human PCK1 knockout A549 cell line (ab301135)
Western blot: Anti-PCK1 antibody [EPR6938] (Anti-PCK1/PEPC antibody [EPR6938] ab133603) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-PCK1/PEPC antibody [EPR6938] ab133603 was shown to bind specifically to PCK1. A band was observed at 67 kDa in wild-type A549 cell lysates with no signal observed at this size in PCK1 knockout cell line. To generate this image, wild-type and PCK1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-PCK1/PEPC antibody [EPR6938] (Anti-PCK1/PEPC antibody [EPR6938] ab133603) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: PCK1 knockout A549 cell lysate at 20 µg
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: HEK-293 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 67 kDa
Next Generation Sequencing - Human PCK1 knockout A549 cell line (ab301135)
1 bp insertion and 35 bp deletion after Met 138 of WT protein
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