Human ECH1 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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ECH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 20 bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
4)-dienoyl-CoA isomerase, 5)-Delta(2, Delta(3, Delta(3 5) Delta(2 4) dienoyl CoA isomerase, Delta(3 5) Delta(2 4) dienoyl CoA isomerase mitochondrial, Delta3 5 delta2 4 dienoyl CoA isomerase, Dienoyl CoA isomerase, ECH1_HUMAN, Enoyl Coenzyme A hydratase 1, Enoyl Coenzyme A hydratase 1 peroxisomal, HPXEL, MS1034, Peroxisomal enoyl CoA hydratase 1, Peroxisomal enoyl coenzyme A hydratase like protein, mitochondrial
- WB
Lab
Western blot - Human ECH1 knockout HEK-293T cell line (AB266748)
Lanes 1-4 : Merged signal (red and green). Green - ab189255 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa.
ab189255 Anti-ECH1 antibody [EPR15449(B)] 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 4° 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 (ab266748)
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 line (AB266748)
Homozygous : 20 bp deletion in exon 2
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 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
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
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
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com