Human PCK2 knockout MCF7 cell line
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PCK2 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 99%.
View Alternative Names
EC 4.1.1.32, GTP mitochondrial precursor, HGNC:8725, Mitochondrial phosphoenolpyruvate carboxykinase 2, OTTHUMP00000164700, PCKGM_HUMAN, PEP carboxykinase, PEPCK, PEPCK deficiency mitochondrial, PEPCK-M, PEPCK2, Phosphoenolpyruvate carboxykinase 2 (mitochondrial), Phosphoenolpyruvate carboxykinase 2 mitochondrial, Phosphoenolpyruvate carboxykinase [GTP], Phosphoenolpyruvate carboxylase, Phosphopyruvate carboxylase, mitochondrial
- NGS
Supplier Data
Next Generation Sequencing - Human PCK2 knockout MCF7 cell line (AB273845)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift : 99%
Reactivity data
Product details
Recommended control: Human wild-type MCF7 cell line (ab271144). 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage duration
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
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
MEM + 10% FBS + 0.01 mg/ml bovine insulin
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
PCK2 contributes to the regulation of glucose homeostasis. It achieves this by being a part of the gluconeogenic pathway working in concert with cytosolic PCK1. PCK2 does not work alone; it integrates into a larger metabolic framework that includes various enzymes essential for converting different substrates into glucose. This enzyme is notably active during fasting states or periods of high energy demands such as intense exercise ensuring a continuous supply of glucose to maintain cellular functions.
Pathways
PCK2 plays an important role in the gluconeogenesis and TCA cycle. The gluconeogenesis pathway allows the liver and kidney to produce glucose from non-carbohydrate substrates which is integral during fasting and low-carbohydrate diet. It is functionally linked with other key enzymes like pyruvate carboxylase in converting pyruvate to glucose and also cooperates with malate dehydrogenase in the TCA cycle. This cooperation ensures efficient energy production and conversion balance in the cell.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
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Product promise
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