AXIN1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
Images
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 2 and Insertion of the selection cassette in exon 2
Recommended products
AXIN1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
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 2 and Insertion of the selection cassette in exon 2
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- AXIN1
- 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 short-term storage conditions
- -196°C
- 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 (ab255448). 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
Axin 1 alternatively known simply as axin is an important protein involved in cell signaling processes with a molecular weight of approximately 94 kDa. Axin 1 serves as a scaffold protein that plays a critical role in various signaling pathways. It shows expression in multiple tissues including the brain liver and kidney which indicates its involvement in various physiological processes. Due to its structural and functional properties axin 1 can be visualized and studied effectively even through techniques such as western blot.
- Biological function summary
Axin 1 functions as a negative regulator within the Wnt signaling pathway by forming a destruction complex with other proteins. This complex is important in the degradation of beta-catenin thereby preventing excessive activation of Wnt target genes. It interacts with several components such as adenomatous polyposis coli (APC) and glycogen synthase kinase 3 beta (GSK3β) to facilitate these functions. The presence of axin 1 maintains the balance of signaling dynamics ensuring that cell proliferation and differentiation proceed correctly.
- Pathways
Axin 1 plays a significant role in the control of the Wnt and JNK signaling pathways. Within the Wnt pathway it partners with proteins such as APC and GSK3β to regulate beta-catenin levels ensuring proper signal transduction. In the JNK pathway axin 1's interactions influence cell response to stress through modulation of signaling cascades. Its involvement in these pathways illustrates how axin 1 acts as a central component in cellular communication and response strategies.
- Associated diseases and disorders
Axin 1 associates with several conditions including colorectal cancer and hepatocellular carcinoma. Dysregulation of axin 1's function or expression can lead to aberrant Wnt signaling which is a pivotal factor in tumorigenesis linked with these cancers. Moreover proteins like beta-catenin and APC which interact with axin 1 also play a role in the progression of these diseases. Understanding axin 1's role provides insights into potential therapeutic strategies that target these pathways for effective treatment approaches.
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2 product images
Sanger Sequencing - Human AXIN1 (Axin 1) knockout HeLa cell line (ab265521)
Allele-2: Insertion of the selection cassette in exon 2.
Sanger Sequencing - Human AXIN1 (Axin 1) knockout HeLa cell line (ab265521)
Allele-1: 1 bp deletion in exon 2.
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