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WWTR1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.

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Images

Western blot - Human WWTR1 knockout HeLa cell line (AB281598), expandable thumbnail
  • Western blot - Human WWTR1 knockout HeLa cell line (AB281598), expandable thumbnail
  • Sanger Sequencing - Human WWTR1 knockout HeLa cell line (AB281598), expandable thumbnail

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

Knockout validation

Sanger Sequencing, Western blot

Alternative names

Recommended products

WWTR1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.

Key facts

Cell type

HeLa

Form

Liquid

Disease

Adenocarcinoma

Concentration
Loading...

Properties

Gene name

WWTR1

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger 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

Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab275466). 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.

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

TAZ also known as WWTR1 (WW Domain-Containing Transcription Regulator 1) acts as a transcriptional co-activator in cells. TAZ has a molecular mass of approximately 45 kDa. This protein appears in various tissues including the lung kidney brain and heart. TAZ interacts with other transcription factors to influence gene expression playing a role in cellular signaling and development.

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.

Associated diseases and disorders

TAZ associates with cancer and fibrosis. In many cancers TAZ exhibits high activity often linked with poor prognosis due to its role in promoting cell proliferation and survival. TAZ interacts with beta-catenin in cancer contexts influencing signaling that drives cancer progression. In fibrosis aberrant TAZ signaling leads to excessive tissue scarring working alongside CTGF (Connective Tissue Growth Factor) to support this pathological development.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Western blot - Human WWTR1 knockout HeLa cell line (ab281598), expandable thumbnail

    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] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-TAZ antibody [CL0371] 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 (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.

    All lanes: Western blot - Anti-TAZ antibody [CL0371] (Anti-TAZ antibody [CL0371] ab242313) 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 3: A549 cell lysate at 20 µg

    Lane 4: K562 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 44 kDa

    Observed band size: 52 kDa

  • Western blot - Human WWTR1 knockout HeLa cell line (ab281598), expandable thumbnail

    Western blot - Human WWTR1 knockout HeLa cell line (ab281598)

    False colour image of Western blot: Anti-TAZ antibody staining at 1 ug/ml shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-TAZ antibody 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 °. 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: Western blot - Anti-TAZ antibody (Anti-TAZ antibody ab110239) 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 3: A549 cell lysate at 20 µg

    Lane 4: K562 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 44 kDa

    Observed band size: 52 kDa

  • Sanger Sequencing - Human WWTR1 knockout HeLa cell line (ab281598), expandable thumbnail

    Sanger Sequencing - Human WWTR1 knockout HeLa cell line (ab281598)

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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