Human HADHA knockout HEK-293T cell lysate
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HADHA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 8 bp deletion in exon 1.
View Alternative Names
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
- WB
Lab
Western blot - Human HADHA knockout HEK-293T cell lysate (AB257464)
Lane 1 : Wild-type HEK-293T cell lysate (20 μg)
Lane 2 : HADHA knockout HEK-293T cell lysate (20 μg)
Lane 3 : HepG2 cell lysate (20 μg)
Lane 4 : SH-SY5Y cell lysate (20 μg)
Lanes 1-4 : Merged signal (red and green). Green - ab200652 observed at 82 kDa. Red - loading control ab8245 observed at 37 kDa.
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 ab257464) was used. Wild-type and HADHA knockout samples were subjected to SDS-PAGE. ab200652 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 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-HADHA antibody [EPR17939] (<a href='/en-us/products/primary-antibodies/hadha-antibody-epr17939-ab200652'>ab200652</a>) 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 2:
Western blot - Human HADHA knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-hadha-knockout-hek-293t-cell-line-ab266274'>ab266274</a>)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
SH-SY5Y cell lysate at 20 µg
Predicted band size: 83 kDa
Observed band size: 82 kDa
false
- WB
Lab
Western blot - Human HADHA knockout HEK-293T cell lysate (AB257464)
Lane 1 : Wild-type HEK-293T cell lysate (20 μg)
Lane 2 : HADHA knockout HEK-293T cell lysate (20 μg)
Lane 3 : HepG2 cell lysate (20 μg)
Lane 4 : SH-SY5Y cell lysate (20 μg)
Lanes 1-4 : Merged signal (red and green). Green - ab203114 observed at 82 kDa. Red - loading control ab8245 observed at 37 kDa.
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 ab257464) was used. Wild-type and HADHA knockout samples were subjected to SDS-PAGE. ab203114 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 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-HADHA antibody [EPR17940] (<a href='/en-us/products/primary-antibodies/hadha-antibody-epr17940-ab203114'>ab203114</a>) 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 2:
Western blot - Human HADHA knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-hadha-knockout-hek-293t-cell-line-ab266274'>ab266274</a>)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
SH-SY5Y cell lysate at 20 µg
Predicted band size: 83 kDa
Observed band size: 82 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human HADHA knockout HEK-293T cell lysate (AB257464)
Allele-1 : 8 bp deletion in exon 1
- Sanger seq
Unknown
Sanger Sequencing - Human HADHA knockout HEK-293T cell lysate (AB257464)
Allele-2 : 1 bp deletion in exon 1
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
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
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
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.
Pathways
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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