CPT2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 2 bp deletion in exon 1.
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Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 2 bp deletion in exon 1
Alternative names
CPT 1, CPT II, CPT2_HUMAN, CPTASE, Carnitine O palmitoyltransferase 2 mitochondrial, Carnitine O-palmitoyltransferase 2, Carnitine palmitoyltransferase 2, Carnitine palmitoyltransferase II, IIAE4, mitochondrial
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CPT2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 2 bp deletion in exon 1.
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 2 bp deletion in exon 1
- Antibiotic resistance
Puromycin 1µg/mL
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
CPT2
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 2 US: 2
- Adherent/suspension
Adherent
- Gender
Female
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.
Storage
- Shipped at conditions
Dry Ice
- Appropriate short-term storage conditions
-196°C
- Appropriate long-term storage conditions
-196°C
Notes
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255928). 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
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Carnitine palmitoyltransferase 2 (CPT2) also known as CPT-2 is an enzyme located mainly in the mitochondria of liver muscle and heart tissues. This enzyme has a molecular mass of approximately 68 kDa. CPT2 plays a mechanical role in the transfer of long-chain fatty acids from the cytosol into the mitochondrial matrix completing the process initiated by CPT1 on the outer mitochondrial membrane. CPT2 acts on the inner mitochondrial membrane converting acylcarnitine back into acyl-CoA which is an essential step in beta-oxidation. Expression of CPT2 is higher in tissues with elevated fatty acid oxidation rates.
- Biological function summary
CPT2 is integral to the mitochondrial pathway of fatty acid metabolism. It functions as part of the carnitine shuttle system a complex important for the conversion of acylcarnitine to acyl-CoA inside the mitochondria. This role is vital for the full oxidation of long-chain fatty acids. Through its action CPT2 aids in maintaining energy homeostasis and producing ketone bodies especially during periods of fasting or strenuous exercise.
- Pathways
CPT2 is involved in both the beta-oxidation and the carnitine shuttle pathways. These pathways facilitate the energy production from fatty acids particularly during states of low carbohydrate availability. CPT2 works closely with CPT1 as both enzymes form the gateway for the entry of long-chain fatty acids into mitochondria. Together with enzymes from the tricarboxylic acid cycle CPT2 helps sustain ATP production under such metabolic conditions.
- Associated diseases and disorders
Defects in CPT2 can lead to disorders like CPT2 deficiency which present with symptoms of muscle weakness and recurrent rhabdomyolysis especially during prolonged fasting or significant exercise. This deficiency can also result in hypoketotic hypoglycemia and liver dysfunction. CPT2-related disorders often involve altered interactions with proteins such as the acyl-CoA dehydrogenases impacting the normal fatty acid oxidation process and causing metabolic disturbances.
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3 product images
Western blot - Human CPT2 knockout HeLa cell line (ab265931)
Lanes 1- 2: Merged signal (red and green). Green - Anti-CPT2 antibody [EPR13626] - C-terminal ab181114 observed at 74 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-CPT2 antibody [EPR13626] - C-terminal ab181114 was shown to react with CPT2/CPT1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265931 (knockout cell lysate Human CPT2 knockout HeLa cell lysate ab257180) was used. Wild-type HeLa and CPT2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-CPT2 antibody [EPR13626] - C-terminal ab181114 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (Anti-CPT2 antibody [EPR13626] - C-terminal ab181114) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CPT2 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 74 kDa
Observed band size: 74 kDa
Sanger Sequencing - Human CPT2 knockout HeLa cell line (ab265931)
Allele-2: 1 bp deletion in exon 1.
Sanger Sequencing - Human CPT2 knockout HeLa cell line (ab265931)
Allele-1: 2 bp deletion in exon 1.
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