WNT5A KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 58 bp deletion, 1bp deletion.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 58 bp deletion, 1bp deletion
Alternative names
Protein Wnt-5a, WNT 5A protein, WNT 5A protein precursor, WNT5A_HUMAN, Wingless type MMTV integration site family member 5A, hWNT 5A
Recommended products
WNT5A KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 58 bp deletion, 1bp deletion.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 58 bp deletion, 1bp deletion
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- WNT5A
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- 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
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
Wnt5a also known simply as Wnt-5a is a member of the Wnt family of signaling proteins. It plays a role in non-canonical Wnt signaling which means it does not involve the typical Wnt/beta-catenin pathway. The protein has a molecular weight of approximately 42 kDa and is widely expressed in various tissues including the brain lungs and reproductive organs. Wnt5a influences several cellular processes such as cell movement polarity and organ development by binding to Frizzled family receptors and co-receptors like ROR1 and ROR2.
- Biological function summary
Wnt5a drives critical processes in development and tissue homeostasis. It acts by guiding cell migration and establishing cell polarity. Wnt5a is not part of a single large complex but interacts with membrane receptors and co-receptors to exert its effects. This interaction initiates signaling cascades that regulate cytoskeletal dynamics which is essential in both embryonic development and adult tissue maintenance. Wnt5a's role is pivotal in maintaining cellular organization and function across different biological contexts.
- Pathways
Wnt5a participates significantly in the planar cell polarity pathway and the Wnt/Ca2+ signaling pathway. In these pathways Wnt5a affects the activity of proteins such as Dishevelled and CaMKII to influence cellular functions independently from beta-catenin. These pathways regulate processes like tissue morphogenesis and cell orientation maintaining the spatial arrangement and structural integrity of tissues.
- Associated diseases and disorders
Wnt5a has been linked to cancer progression and inflammatory diseases such as rheumatoid arthritis. It interacts with proteins like ROR1 and ROR2 during these pathologies. In cancer abnormal Wnt5a signaling can lead to enhanced cell migration and invasion contributing to metastasis. In rheumatoid arthritis Wnt5a may drive inflammatory pathways and joint tissue damage. Researchers consider Wnt5a a target for therapeutic interventions in these conditions with ongoing studies focused on modulating its activity for disease management.
Product promise
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2 product images
Next Generation Sequencing - Human WNT5A knockout HeLa cell line (ab264019)
7 bp deletion after Gln106 (allele 1) and 4 bp deletion after Phe107 (allele 2) of the WT protein
Western blot - Human WNT5A knockout HeLa cell line (ab264019)
False colour image of Western blot: Anti-WNT5A antibody staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to WNT5A. A band was observed at 40-45 kDa in wild-type HeLa cell lysates with no signal observed at this size in WNT5A knockout cell line ab264019 (knockout cell lysate Human WNT5A knockout HeLa cell lysate ab264515). To generate this image, wild-type and WNT5A knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Anti-WNT5A antibody at 1/1000 dilution
Lane 1: Wild-type HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: Western blot - Human WNT5A knockout HeLa cell line (ab264019)
Lane 2: Western blot - Human WNT5A knockout HeLa cell lysate (Human WNT5A knockout HeLa cell lysate ab264515) at 20 µg
Lane 3: U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 20 µg
Lane 4: SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg
SecondaryLanes 1 - 4: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution
Lanes 1 - 4: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Predicted band size: 42 kDa
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