ECE1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 131 bp deletion in exon 4.
Images
Key facts
- Cell type
- U-87 MG
- Species or organism
- Human
- Tissue
- Brain
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 131 bp deletion in exon 4.
Alternative names
ECE, ECE1_HUMAN, Endothelin-converting enzyme 1
Recommended products
ECE1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 131 bp deletion in exon 4.
Key facts
- Cell type
- U-87 MG
- Species or organism
- Human
- Tissue
- Brain
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 131 bp deletion in exon 4.
- Disease
- Glioblastoma
- Concentration
Properties
- Gene name
- ECE1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- Zygosity
- Homozygous
Quality control
- STR analysis
- VWA, CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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
- EMEM + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The ECE1 gene encodes for endothelin-converting enzyme 1 also referred to as ECE-1. This enzyme weighs approximately 120 kDa and functions mechanically to convert big endothelins into active endothelins. ECE-1 is present in many tissues with higher expression in the endothelium kidneys and lungs. Through its metalloprotease activity ECE-1 plays a central role in regulating endothelin peptide activation which is important for vasoconstriction and other physiological processes.
- Biological function summary
ECE-1 takes part in maintaining vascular homeostasis and blood pressure regulation. It does not function as part of a complex but exerts its effects through the cleavage of specific precursor proteins such as big endothelins. By controlling the conversion of these precursors to active peptidic forms ECE-1 directly influences the vascular tone and endothelial function.
- Pathways
ECE-1 impacts the endothelin signaling pathway significantly where it plays a role in converting big endothelin-1 to a potent vasoconstrictor endothelin-1. This pathway involves other proteins like endothelin receptors which mediate the physiological effects of endothelins on smooth muscles and vascular tissues. Additionally ECE-1 interacts with the renin-angiotensin-aldosterone system (RAAS) linking it to blood pressure modulation and sodium balance.
- Associated diseases and disorders
ECE-1 is associated with conditions such as hypertension and pulmonary arterial hypertension (PAH). Elevated activity or expression levels can contribute to increased vascular resistance leading to heightened blood pressure. Through its role in endothelin signaling ECE-1 also links to proteins like endothelin-1 which are implicated in the pathophysiology of these cardiovascular disorders.
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2 product images
Western blot - Human ECE1 knockout U-87 MG cell line (ab288712)
False colour image of Western blot: Anti-ECE1 antibody staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to ECE1. A band was observed at 135 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in ECE1 knockout cell line ab288712 (knockout cell lysate ab289605). To generate this image, wild-type and ECE1 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Anti-ECE1 antibody
Lane 1: Wild-type U-87 MG cell lysate at 20 µg
Lane 2: ECE1 knockout U-87 MG cell lysate at 20 µg
Lane 3: NIH/3T3 cell lysate at 20 µg
Lane 4: HUVEC cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 87 kDa
Observed band size: 135 kDa
Sanger Sequencing - Human ECE1 knockout U-87 MG cell line (ab288712)
131 bp deletion in exon 4.
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