Human WNT5A knockout HeLa cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
WNT5A KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 58 bp deletion, 1bp deletion. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
Protein Wnt-5a, WNT 5A protein, WNT 5A protein precursor, WNT5A_HUMAN, Wingless type MMTV integration site family member 5A, hWNT 5A
- WB
Lab
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] (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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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 (<a href='/en-us/products/cell-lysates/human-wnt5a-knockout-hela-cell-lysate-ab264515'>ab264515</a>) 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
Secondary
Lanes 1 - 4:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Lanes 1 - 4:
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 1/20000 dilution
Predicted band size: 42 kDa
false
- NGS
Supplier Data
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
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
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
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