Human CPT1A knockout HEK-293T cell line
- Advanced Validation
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CPT1A KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 3 and Insertion of the selection cassette in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
CPT 1, CPT I, CPT1-L, CPT1A_HUMAN, CPTI-L, Carnitine O palmitoyltransferase 1 liver isoform, Carnitine O palmitoyltransferase I liver isoform, Carnitine O-palmitoyltransferase 1, Carnitine O-palmitoyltransferase I, Carnitine palmitoyltransferase 1A, Carnitine palmitoyltransferase 1A (liver), Carnitine palmitoyltransferase I, Carnitine palmitoyltransferase I liver, L CPT1, liver isoform
- WB
Lab
Western blot - Human CPT1A knockout HEK-293T cell line (AB266319)
Lanes 1- 2 : Merged signal (red and green). Green - ab220789 observed at 88 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab220789 was shown to react with CPT1A in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266319 (knockout cell lysate ab256880) was used. Wild-type HEK-293T and CPT1A knockout HEK-293T 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. ab220789 and 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 (ab216773) and 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-CPT1A antibody [EPR21843-71-1C] (<a href='/en-us/products/primary-antibodies/cpt1a-antibody-epr21843-71-1c-ab220789'>ab220789</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
CPT1A knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human CPT1A knockout HEK-293T cell line (ab266319)
Predicted band size: 88 kDa
Observed band size: 88 kDa
false
- WB
Lab
Western blot - Human CPT1A knockout HEK-293T cell line (AB266319)
Lanes 1- 2 : Merged signal (red and green). Green - ab234111 observed at 88 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab234111 was shown to react with CPT1A in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266319 (knockout cell lysate ab256880) was used. Wild-type HEK-293T and CPT1A knockout HEK-293T 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. ab234111 and 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 (ab216773) and 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-CPT1A antibody [EPR21843-71-2F] (<a href='/en-us/products/primary-antibodies/cpt1a-antibody-epr21843-71-2f-ab234111'>ab234111</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
CPT1A knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human CPT1A knockout HEK-293T cell line (ab266319)
Predicted band size: 88 kDa
Observed band size: 88 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CPT1A knockout HEK-293T cell line (AB266319)
Allele-1 : 17 bp deletion in exon 3
- Sanger seq
Unknown
Sanger Sequencing - Human CPT1A knockout HEK-293T cell line (AB266319)
Allele-2 : Insertion of the selection cassette in exon 3.
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
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
CPT1A plays a vital role in energy production by facilitating mitochondrial fatty acid oxidation. It does not function as a solitary enzyme but often associates with other enzymes forming a multiprotein complex that includes acyl-CoA synthetase. This complex enables efficient fatty acid transport and oxidation converting stored fats into usable energy. As CPT1A primarily operates in the liver its activity significantly impacts overall lipid and energy homeostasis demonstrating its critical regulatory role.
Pathways
CPT1A is central to the fatty acid beta-oxidation pathway an important process for breaking down fatty acids to produce energy. This pathway also involves proteins such as CPT1B a related isoform present in muscle tissues. CPT1A's function is important in the liver's capacity to regulate energy balance and respond to metabolic demands. Additionally CPT1A interacts within the signaling pathway of AMPK (AMP-activated protein kinase) which further integrates it into broader metabolic regulation networks linking energy status to cellular function.
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
Complementary Products
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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