ACVRL1 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 5 and 2 bp deletion in exon 5.
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Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 5 and 2 bp deletion in exon 5
Recommended products
ACVRL1 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 5 and 2 bp deletion in exon 5.
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 5 and 2 bp deletion in exon 5
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
ACVRL1
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
Cell culture
- Biosafety level
EU: 2 US: 2
- Viability
~ 80%
- Adherent/suspension
Adherent
- Gender
Female
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.
Storage
- Shipped at conditions
Dry Ice
- Appropriate long-term storage conditions
-196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab271142). 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM (High Glucose) + 10% FBS
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 2x10E4 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 2x10E4 cells/cm2 is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.
• Do not allow the cell density to exceed 7x10E4 cells/cm2.
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
- Activity summary
ALK-1 also known as ACVRL1 or ALK1 protein is a receptor serine/threonine kinase with a molecular mass of approximately 55 kDa. It plays an important role in the TGF-beta superfamily signaling particularly in angiogenesis and blood vessel development. Expressed mainly in endothelial cells ALK-1 mediates cellular responses essential for maintaining vascular stability. The careful balance of its signaling is necessary indicating its importance in physiological conditions.
- Biological function summary
The function of ALK-1 extends to the formation and maintenance of vascular endothelium as part of the larger receptor complex. It interacts with ligands such as BMP9 and BMP10 which regulate endothelial cell proliferation and differentiation. ALK-1's role as part of this receptor complex highlights its involvement in controlling the fine balance between angiogenic and anti-angiogenic processes. Its biological activity is vital for healthy vascular development and homeostasis.
- Pathways
ALK-1 activation is a part of the BMP signaling pathway a branch of the larger TGF-beta pathway. This pathway is significant in various cellular processes including growth and differentiation. ALK-1 interacts with other signalling mediators like SMAD1/5/8 proteins which translocate to the nucleus to affect gene expression. Connecting with these pathways ALK-1 aligns closely with ACVR2A and ENG (Endoglin) which further highlight its role in vascular biology.
- Associated diseases and disorders
ALK-1 mutations or dysregulation leads to conditions such as hereditary hemorrhagic telangiectasia (HHT) and pulmonary arterial hypertension (PAH). These conditions showcase the important role ALK-1 plays in vascular integrity and signaling. ALK-1 is connected to disorders through interactions with proteins such as ENG further highlighting its involvement in vascular-related pathologies. Maintaining precise ALK-1 function is pivotal to preventing these disorders indicating its potential as a therapeutic target or biomarker.
Product promise
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
2 product images
Sanger Sequencing - Human ACVRL1 (ALK-1) knockout HeLa cell line (ab265547)
Allele-1: 2 bp deletion in exon 5.
Sanger Sequencing - Human ACVRL1 (ALK-1) knockout HeLa cell line (ab265547)
Allele-2: 1 bp deletion in exon 5.
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