Human WNT5A knockout HeLa cell lysate
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- WB
Lab
Western blot - Human WNT5A knockout HeLa cell lysate (AB264515)
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 (<a href='/en-us/products/cell-lines/human-wnt5a-knockout-hela-cell-line-ab264019'>ab264019</a>)
Lane 2:
Western blot - 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
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
Complementary Products
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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