Human ECH1 knockout HEK-293T cell lysate
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- WB
Lab
Western blot - Human ECH1 knockout HEK-293T cell lysate (AB257933)
Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : ECH1 knockout HEK293T cell lysate (20 ug)
Lane 3 : A549 cell lysate (20 ug)
Lane 4 : U-2 OS cell lysate (20 ug)
ab189255 was shown to specifically react with ECH1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266748 (knockout cell lysate ab257933) was used. Wild-type and ECH1 knockout samples were subjected to SDS-PAGE. ab189255 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4oC at 1 in 1000 dilution 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-ECH1 antibody [EPR15449(B)] - C-terminal (<a href='/en-us/products/primary-antibodies/ech1-antibody-epr15449b-c-terminal-ab189255'>ab189255</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ECH1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ECH1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ech1-knockout-hek-293t-cell-line-ab266748'>ab266748</a>)
Lane 3:
A549 cell lysate at 20 µg
Lane 4:
U-2 OS cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 36 kDa,80 kDa
Observed band size: 100 kDa,36 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ECH1 knockout HEK-293T cell lysate (AB257933)
Homozygous : 20 bp deletion in exon 2
Complementary Products
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
Zygosity
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
The ECH1 protein facilitates the breakdown and utilization of unsaturated fatty acids thereby providing energy for cellular processes. It acts independently and is not a part of a large protein complex. Mitochondria rely on ECH1 to handle specific intermediates in fatty acid metabolism which impacts how cells manage their energy resources. This makes ECH1 essential for maintaining energy homeostasis particularly during periods of increased metabolic demand.
Pathways
ECH1 is integral to the mitochondrial fatty acid beta-oxidation pathway. This pathway is important for converting fatty acids into acetyl-CoA units which then enter the Krebs cycle for further energy production. ECH1 interacts with other proteins like ACAA2 which contributes to the final step of fatty acid degradation. Alongside this it aligns with the peroxisomal biogenesis pathway influencing the balance between metabolic processes within mitochondria and peroxisomes.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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