Human WWTR1 knockout HeLa cell line
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WWTR1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. 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
DKFZP586I1419, FLJ27004, FLJ45718, OTTHUMP00000215994, OTTHUMP00000215995, OTTHUMP00000215996, OTTHUMP00000216001, Transcriptional coactivator with PDZ-binding motif, WW domain containing transcription regulator 1, WW domain-containing transcription regulator protein 1, WWTR1_HUMAN
- WB
Lab
Western blot - Human WWTR1 knockout HeLa cell line (AB281598)
Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : WWTR1 knockout HeLa cell lysate 20 μg
Lane 3 : A549 cell lysate 20 μg
Lane 4 : K562 cell lysate 20 μg
False colour image of Western blot : Anti-TAZ antibody staining at 1 μg/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab110239 was shown to bind specifically to TAZ. A band was observed at 52 kDa in wild-type HeLa cell lysates with no signal observed at this size in WWTR1 knockout cell line ab281598 (knockout cell lysate ab282950). To generate this image, wild-type and WWTR1 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:
Western blot - Anti-TAZ antibody (<a href='/en-us/products/primary-antibodies/taz-antibody-ab110239'>ab110239</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human WWTR1 knockout HeLa cell lysate (ab282950) at 20 µg
Lane 3:
A549 cell lysate at 20 µg
Lane 4:
K562 cell lysate at 20 µg
Secondary
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
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
Predicted band size: 44 kDa
Observed band size: 52 kDa,37 kDa
false
- WB
Lab
Western blot - Human WWTR1 knockout HeLa cell line (AB281598)
False colour image of Western blot : Anti-TAZ antibody [CL0371] staining at 1 ug/ml shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution shown in red. In Western blot ab242313 was shown to bind specifically to TAZ. A band was observed at 52 kDa in wild-type HeLa cell lysates with no signal observed at this size in WWTR1 knockout cell line ab281598 (knockout cell lysate ab282950). To generate this image wild-type and WWTR1 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 °. 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
All lanes:
Western blot - Anti-TAZ antibody [CL0371] (<a href='/en-us/products/primary-antibodies/taz-antibody-cl0371-ab242313'>ab242313</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
WWTR1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human WWTR1 knockout HeLa cell line (ab281598)
Lane 3:
A549 cell lysate at 20 µg
Lane 4:
K562 cell lysate at 20 µg
Predicted band size: 44 kDa
Observed band size: 52 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human WWTR1 knockout HeLa cell line (AB281598)
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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
TAZ works as a part of the Hippo signaling pathway regulating cell growth proliferation and apoptosis. It changes cellular responses through interactions with other proteins such as TEAD transcription factors. TAZ can act within the cytoplasm or the nucleus changing its location in the cell based on physiological signals. It operates independently as well as in concert with other molecules assisting in maintaining tissue homeostasis and regeneration.
Pathways
TAZ functions as an essential element of the Hippo pathway which controls organ size and suppresses cancer. It plays significant roles in the PI3K-AKT signaling pathway coordinating with YAP (Yes-associated protein) a well-known partner in these pathways. TAZ and YAP together influence transcriptional outcomes for numerous genes adjusting to the needs of various cellular contexts which affect cell behavior significantly.
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
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Advanced Validation
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Primary Antibodies
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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