HADHA 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 8 bp deletion in exon 1.
HEK-293T
Human
Kidney
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 8 bp deletion in exon 1
3 ketoacyl Coenzyme A (CoA) thiolase alpha subunit, 3 oxoacyl CoA thiolase, 78 kDa gastrin-binding protein, ECHA_HUMAN, HADHA, Hydroxyacyl Coenzyme A dehydrogenase/3 ketoacyl Coenzyme A thiolase/enoyl Coenzyme A hydratase (trifunctional protein) alpha subunit, LCEH, LCHAD, Long chain 3-hydroxyacyl-CoA dehydrogenase, MTPA, Mitochondrial long chain 2 enoyl Coenzyme A (CoA) hydratase alpha subunit, Mitochondrial long chain L 3 hydroxyacyl Coenzyme A dehydrogenase alpha subunit, Mitochondrial trifunctional enzyme alpha subunit, Mitochondrial trifunctional protein alpha subunit, TP-alpha, Thiolase/enoyl Coenzyme A hydratase (trifunctional protein) alpha subunit, Trifunctional enzyme subunit alpha mitochondrial precursor
HADHA 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 8 bp deletion in exon 1.
HEK-293T
Human
Kidney
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 8 bp deletion in exon 1
HADHA
Knockout
CRISPR technology
Sanger Sequencing, Western blot
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
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.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T cell line ab255449). 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.
This supplementary information is collated from multiple sources and compiled automatically.
The HADHA protein also called hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha is an essential component of the mitochondrial trifunctional protein complex. This protein has a molecular mass of about 79 kDa and is expressed mainly in tissues with high fatty acid oxidation rates like liver heart and muscle. HADHA plays a significant role in the beta-oxidation of long-chain fatty acids acting on hydroxyacyl-CoA substrates during this critical metabolic process.
HADHA is a part of the mitochondrial trifunctional protein complex which consists of four alpha and four beta subunits. It facilitates the hydration of enoyl-CoA to 3-hydroxyacyl-CoA and the subsequent dehydrogenation to 3-ketoacyl-CoA. This enzyme works closely with its partner the HADHB protein to carry out these reactions efficiently. These functions are important for energy production as they are steps in the breakdown of fatty acids necessary for ATP generation.
HADHA participates in the mitochondrial beta-oxidation pathway an essential pathway for energy production from fats. Alongside HADHB it catalyzes key reactions that allow the progressive shortening of fatty acid chains which further feeds into the citric acid cycle. This pathway links HADHA not only to HADHB but also to other enzymes involved in lipid metabolism and energy homeostasis including medium-chain specific acyl-CoA dehydrogenase (MCAD) reflecting its role in comprehensive metabolic networks.
Defects in HADHA are associated with mitochondrial trifunctional protein deficiency and long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency. Both disorders disrupt normal fatty acid oxidation leading to a spectrum of symptoms including hypoketotic hypoglycemia and cardiomyopathy. These conditions highlight the relationship between HADHA and other proteins involved in fatty acid metabolism such as HADHB further highlighting their collective role in maintaining cellular energy balance.
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Lanes 1-4: Merged signal (red and green). Green - Anti-HADHA antibody [EPR17939] ab200652 observed at 82 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-HADHA antibody [EPR17939] ab200652 Anti-HADHA antibody [EPR17939] was shown to specifically react with HADHA in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266274 (knockout cell lysate Human HADHA knockout HEK-293T cell lysate ab257464) was used. Wild-type and HADHA knockout samples were subjected to SDS-PAGE. Anti-HADHA antibody [EPR17939] ab200652 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° at 1 in 1000 and 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-HADHA antibody [EPR17939] (Anti-HADHA antibody [EPR17939] ab200652) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HADHA knockout HEK-293T cell lysate at 20 µg
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: SH-SY5Y cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 82 kDa
Lanes 1-4: Merged signal (red and green). Green - Anti-HADHA antibody [EPR17940] ab203114 observed at 82 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-HADHA antibody [EPR17940] ab203114 Anti-HADHA antibody [EPR17940] was shown to specifically react with HADHA in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266274 (knockout cell lysate Human HADHA knockout HEK-293T cell lysate ab257464) was used. Wild-type and HADHA knockout samples were subjected to SDS-PAGE. Anti-HADHA antibody [EPR17940] ab203114 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° at 1 in 1000 and 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-HADHA antibody [EPR17940] (Anti-HADHA antibody [EPR17940] ab203114) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HADHA knockout HEK-293T cell lysate at 20 µg
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: SH-SY5Y cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 82 kDa
Allele-1: 8 bp deletion in exon 1
Allele-2: 1 bp deletion in exon 1.
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